KR20150139955A - 2-acetylnaphtho[2,3-b]furan-4,9-dione for use on treating cancer - Google Patents

Abstract

The present invention relates to a naphthofuran compound, a polymorph of a naphthofuran compound, a naphthofuran compound in the form of a particle, a purified composition containing at least one naphthofuran compound, at least one naphthofuran compound in the form of a particle, And methods of using these naphthofuran compounds, polymorphs, purified compositions and / or particle forms to treat a subject in need of treatment.

Description

2-ACETYLNAPHTHO [2,3-B] FURAN-4,9-DIONE FOR USE ON TREATING CANCER}

Related application

This application is related to U.S. Provisional Application No. 61 / 810,117, filed April 9, 2013; U.S. Provisional Application No. 61 / 830,068, filed June 1, 2013; U.S. Provisional Application No. 61 / 932,179, filed January 27, 2014; And U.S. Provisional Application No. 61 / 938,386, filed February 11, 2014, as priority claims. Each of which is incorporated herein by reference in its entirety.

Field of invention

The present invention relates to a naphthofuran compound, a polymorph of a naphthofuran compound, a naphthofuran compound in the form of a particle, a purified composition containing at least one naphthofuran compound, at least one naphthofuran compound in the form of a particle, And methods of using these naphthofuran compounds, polymorphs, purified compositions and / or particle forms to treat a subject in need of treatment.

In the United States alone, the cancer mortality rate reaches hundreds of thousands of years. Despite advances in the treatment of certain forms of cancer through surgery, radiation therapy and chemotherapy, various types of cancer are inherently untreatable. Although effective treatments for certain cancers are available, the side effects of such treatments can become serious and greatly diminish the quality of life.

The most common chemotherapeutic agents have toxicity and limited efficacy, especially in patients with advanced solid tumors. The chemotherapeutic agent destroys cancer cells as well as non-cancer cells. The therapeutic index of such compounds (a measure of the ability of a therapy to distinguish between cancer cells and normal cells) can be very low. Often, doses of chemotherapeutic drugs effective in killing cancer cells will also kill normal cells, particularly normal cells (e.g., epithelial cells), which often undergo cell division. When normal cells are affected by therapy, hair loss, inhibition of hematopoiesis, and nausea can occur. Depending on the general health of the patient, the side effects may interfere with the administration of the chemotherapy, or at least be very uncomfortable to the patient, uncomfortable, and may significantly impair the quality of life of the remaining patient. Even in the case of cancer patients responding to chemotherapy with tumor regression, the tumor response often does not involve prolongation of progression-free survival (PFS) or prolongation of overall survival (OS). In fact, cancer often progresses rapidly after the initial response to chemotherapy, forming more metastases. The recurrent cancer becomes resistant or refractory to the chemotherapeutic agent. This rapid recurrence and refinement after chemotherapy is considered to be caused by cancer stem cells.

Recent studies have revealed the presence of cancer stem cells (CSC, also referred to as tumor-initiated cells or cancer stem-like cells), which have self-renewal potential and are considered to be the primary cause of malignancy, recurrence and metastasis. Importantly, CSCs have inherent resistance to conventional therapies. Thus, targeting agents with activity against cancer stem cells have great potential for cancer patients ( J Clin Oncol . Jun 10; 26 (17)). Therefore, conventional chemotherapy can kill large cancer cells, but they leave cancer stem cells. Cancer stem cells can grow more rapidly after reduction of non-stem cell cancer cells by chemotherapy, which is considered to be a mechanism for rapid recurrence after chemotherapy.

Accordingly, there is a need for the discovery of compounds and pharmaceutical compositions for selectively targeting cancer cells, compounds and pharmaceutical compositions for targeting cancer stem cells, methods for the preparation of these compounds, pharmaceutical compositions for clinical application, There is a need for a method of administering the compound to a subject.

The literature cited herein is not to be construed as prior art to the claimed invention.

In co-owned PCT applications, published as WO 2009/036099, WO 2009/036101 and WO 2011/116399, the contents of both of which are incorporated herein by reference, the disclosures relate to novel naphthofuran compounds, naphthofuran compounds , A purified composition containing at least one naphthofuran compound and a naphthofuran compound in the form of particles. These naphthofuran compounds (including those in particle form), polymorphs and purified compositions are selective inhibitors of cancer stem cells and STAT3. WO 2009/036099 and WO 2009/036101 disclose that naphthofuran compounds target cancer stem cells. They also inhibit STAT3 and suppress non-stem cancer cells. These compounds can kill many different types of cancer cells without destroying the normal cells under specific exposure conditions. Therefore, the compounds may be used for the treatment of cancer, especially for the treatment and prevention of intractable, recurrent, metastatic or STAT3-expressing cancer. These publications describe methods for preparing pharmaceutical compositions of naphthofuran compounds, derivatives and intermediates thereof and related compounds.

The present invention provides novel formulation and use of these naphthofuran compounds (including those in particulate form), polymorphs and purified compositions in a variety of directions, including treatment of cell proliferative disorders, delayed progression, prevention of recurrence, ≪ / RTI > For example, naphthofuran compounds (including those in particulate form), polymorphs and purified compositions are useful for treating cancer, delaying progression, preventing recurrence, alleviating or otherwise alleviating symptoms. In some embodiments, the cancer is selected from the group consisting of esophageal cancer, gastric adenocarcinoma, glioma adenocarcinoma, chondrosarcoma, colon cancer (colorectal cancer), colon adenocarcinoma, rectal adenocarcinoma, colon adenocarcinoma (colon adenocarcinoma), breast cancer, ovarian cancer, head and neck cancer, Melanoma, gastric adenocarcinoma, and adrenocortical carcinoma. In some embodiments, the cancer is an esophageal cancer. In some embodiments, the cancer is a gastric cancer. In some embodiments, the cancer is a gastric adenocarcinoma. In some embodiments, the cancer is refractory. In some embodiments, the cancer is recurrent. In some embodiments, the cancer is metastatic. In some embodiments, the cancer is associated with overexpression of STAT3.

The method according to the invention for the treatment of cancer (or neoplasia) in a human, mammalian or animal subject, delaying progression, preventing recurrence, preventing relapse, inhibiting metastasis, alleviating symptoms and / Lt; RTI ID = 0.0 > and / or < / RTI > pharmaceutical composition of the present invention. For example, anti-neoplastic activity may be an anticancer activity. For example, the anti-neoplastic activity may include a delay in the volume growth of the neoplasm, a halt in the volume growth of the neoplasm, or a volume reduction in the neoplasm. Neoplasms can include solid tumors, malignant, metastatic, cancer stem cells. Neoplasms include carcinoma, sarcoma, adenocarcinoma, lymphoma or hematologic malignancies. The neoplasm may be refractory to treatment by chemotherapy, radiation therapy and / or hormone therapy. The compounds, products and / or pharmaceutical compositions may be administered to prevent recurrence of the neoplasm. The compounds, products and / or pharmaceutical compositions may be administered as adjunctive therapy for surgical resection. The compounds, products and / or pharmaceutical compositions may be administered, for example, orally and / or intravenously. In some embodiments, the pharmaceutical composition comprises at least (i) a surfactant comprising sodium lauryl sulfate (SLS) or sodium dodecyl sulfate (SDS); (ii) Gelucire (lauroyl polyoxyl glyceride); And Labrafil (linoleoylpolyoxylglyceride). ≪ / RTI >

As used herein, the term "treatment of cancer" may include delaying the progression of cancer (or neoplasm), preventing recurrence, relapse, inhibiting metastasis, alleviating symptoms and / or otherwise improving.

In some embodiments, the pharmaceutical composition comprises at least (i) a surfactant comprising sodium lauryl sulfate (SLS) or sodium dodecyl sulfate (SDS); (ii) gel lucer (lauroyl polyoxyl glyceride); And (iii) labrafil (linoleoylpolyoxyl glyceride).

In some embodiments, the pharmaceutical composition comprises about 27.18 wt% active ingredient, about 0.27 wt% surfactant, about 14.51 wt% gel lucer, and about 58.04 wt% labrafil. In some embodiments, the pharmaceutical composition comprises about 125 mg of active ingredient, about 1.2 mg of a surfactant, about 66.8 mg of gel lucer and about 267 mg of labrafil. In some embodiments, the pharmaceutical composition comprises about 80 mg of active ingredient, about 0.8 mg of a surfactant, about 42.7 mg of gel lucer and about 170.9 mg of labrafil. In some embodiments, the pharmaceutical composition is contained within a capsule. In some embodiments, the capsule has a size of 1 or less.

The methods according to the invention also include the treatment of a disease or disorder in a human, mammal or animal subject suffering from the disease or disorder, delaying the progression, preventing recurrence, alleviating or otherwise alleviating the symptoms. In some embodiments, the disease or disorder is any cancer (or neoplasm) described herein. In some embodiments, the cancer is selected from the group consisting of esophageal cancer, gastric adenocarcinoma, glioma adenocarcinoma, chondrosarcoma, colon cancer, colon adenocarcinoma, rectal adenocarcinoma, colon adenocarcinoma, breast cancer, ovarian cancer, head and neck cancer, melanoma, adenocarcinoma and adrenocortical carcinoma ≪ / RTI >

In some embodiments, the method also includes detecting the level of phosphorylated STAT3 (p-STAT3) in the patient's tissue, wherein the level of p-STAT3 is used as a biomarker for patient selection. In some embodiments, the tissue phosphorylated STAT3 level is greater than the benchmark level (more than 10% tumor cells with an intermediate level of p-STAT3). In some embodiments, the cancer is associated with beta -catenin localization in the cell nucleus, not in the cell membrane. In some embodiments, the method comprises detecting a site of beta -catenin expression in a patient's tissue, wherein the site of beta -catenin expression is used as a biomarker for patient selection. In some embodiments, significant beta -catenin expression is detected in the nucleus. In some embodiments, moderate to strong expression of beta -catenin is detected in 20% or more tumor cells.

The administration of the compound, product and / or pharmaceutical composition to a patient suffering from a disease or disorder is considered successful if any various laboratory or clinical outcome is achieved. For example, administration is considered successful if at least one of the symptoms associated with the disease or disorder is alleviated, reduced, inhibited, or does not progress to an additional, i.e., exacerbated, condition. Administration is considered successful if the disorder, e.g., cancer or neoplasm, is showing progress or not progressing further, i.e., in a deteriorated state.

In some embodiments, the compounds, products and / or pharmaceutical compositions described herein are administered in combination with any of the various known therapeutic agents, including, for example, chemotherapeutic agents and other anti-neoplastic agents, antiinflammatory compounds, and / do. In some embodiments, the compounds, products and / or pharmaceutical compositions described herein are useful with, as non-limiting examples, surgical treatment and methods, radiation therapy, chemotherapy and / or hormone or other endocrine-related therapies.

These "combination therapies" may be administered sequentially or concurrently. The compounds, products and / or pharmaceutical compositions described herein and the second therapy may be administered to a subject, preferably a human subject, in the same pharmaceutical composition. Alternatively, the compounds, products and / or pharmaceutical compositions described herein and the second therapy may be administered simultaneously, separately or sequentially to the subject in separate pharmaceutical compositions. The compounds, products and / or pharmaceutical compositions described herein and the second therapy may be administered to a subject by administration of the same or different routes. The compounds, products and / or pharmaceutical compositions described herein may be first administered to a subject, and then a second therapy may be administered to the subject. A second therapy may be administered to the subject first, and then the compounds, products and / or pharmaceutical compositions described herein may be administered to the subject. In some embodiments, the combination therapy of the invention comprises an effective amount of a compound, product, and / or pharmaceutical composition described herein and an effective amount of at least one effective amount of a compound, product, and / or pharmaceutical composition described herein, Other therapies (such as prophylactic or therapeutic agents). In some embodiments, the combination therapies of the present invention work together to provide an additive or synergistic effect by improving the prophylactic or therapeutic effects of the compounds, products and / or pharmaceutical compositions described herein and of the second therapy. In certain embodiments, the combination therapy of the present invention reduces the side effects associated with the second therapy (e.g., prophylactic or therapeutic agents).

In some embodiments, the disease or disorder can be treated by administering a compound, product, and / or pharmaceutical composition as described below. The molar concentration in the blood of the compound is at least about the effective period, at least the effective concentration during the first continuous period shorter than the harmful period, and may be lower than the harmful concentration. The molar concentration in blood may be lower than the effective concentration after the first continuous period. For example, the effective concentration may be about 0.1 μM, about 0.2 μM, about 0.5 μM, about 1 μM, about 2 μM, about 3 μM, about 4 μM, about 5 μM, about 6 μM, about 10 μM, It may be another concentration determined to be valid. For example, the hazardous concentration may be about 1 μM, about 3 μM, about 10 μM, about 15 μM, about 30 μM, about 100 μM or another concentration determined to be detrimental to the skilled artisan. For example, the shelf life may be about 1 hour, 2 hours, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 24 hours, or a period of time determined to be effective by those skilled in the art. For example, the duration of the hazard may be about 12 hours, about 24 hours, about 48 hours, about 72 hours, about 144 hours, or a period of time determined to be detrimental to those skilled in the art.

In some embodiments, a therapeutically effective amount of a compound, product, and / or pharmaceutical composition is greater than the IC ₅₀ of a tumor cell and is selected to produce a blood concentration lower than the IC ₅₀ of a normal cell. In some embodiments, the therapeutically effective amount is high enough to kill the cells of the tumor and is selected to produce a blood concentration lower than the IC ₅₀ of normal cells.

In some embodiments, the compounds, products and / or pharmaceutical compositions may be formulated for administration, for example as tablets, pills, capsules (hard or soft), dragees, powders, granules, suspensions, liquids, gels, , Lozenges, syrups, elixirs, emulsions, oil-in-water emulsions, water-in-oil emulsions and / or potions.

In various embodiments of combination therapy, the compounds of the invention are administered to a patient in a total daily dosage within the range of about 400 mg to about 1,000 mg. In some embodiments, the compounds of the invention are administered to a patient in a total daily dosage within the range of from about 800 mg to about 1,000 mg, preferably twice daily, for example, in the range of about 480 mg BID. The interval between doses may be in the range of about 4 hours to about 16 hours, such as about 12 hours.

In some embodiments, the dose of the compound of the present invention may be varied such that the total daily dose is reduced to a total of 400 to 800 mg per day. In some embodiments, the dosage can be varied such that the total daily dose is reduced to a total of 50 mg to 400 mg per day. In some embodiments, the compounds of the invention may also be taken once a day. In some embodiments, when taken once a day, the interval between doses may be 18 to 30 hours (e.g., about 24 hours). In some embodiments, the compounds of the present invention may also be administered three times daily at a total dose of about 240 to 1,000 mg. When administered three times daily, the time between doses may be from about 4 hours to 8 hours.

In one aspect of the invention, the naphthofuran compounds of the present invention may be used in combination with an anti-mitotic agent, or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, which has been shown to be particularly effective chemotherapeutic agents. Examples of antimitotic agents that may be useful as combination therapy with the compounds of the present invention include paclitaxel (ablucan / taxol), docetaxel (taxotere), BMS-275183, xyotax, tocosal, vinorelbine, , Vinblastine, vindesine, binzolidine, etoposide (VP-16), tenniposide (VM-26), ixabepilone, laurotaxel, But is not limited thereto.

In some embodiments, the second drug used in combination therapy with a compound of the present invention is paclitaxel (ablucan / taxol) or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof. In some embodiments, the paclitaxel is administered to the subject in a total weekly dosage within the range of about 40 mg / m 2 to about 100 mg / m 2. In some embodiments, the paclitaxel is administered to the subject in a total weekly dose of about 80 mg / m 2. In some embodiments, paclitaxel is administered to a subject by IV. In some embodiments, paclitaxel is administered once per week for three weeks per week, i. E., Three weeks, with one week off.

In some embodiments, a compound of the invention may be first administered to a subject, and then paclitaxel may be administered to the subject. Paclitaxel may first be administered to the subject, and then the compound of the invention may be administered to the subject. In such cases, some spacing between administration of the compounds of the present invention and paclitaxel may be included. In some embodiments, the invention encompasses administering a dose of a compound of the invention and a dose of paclitaxel to a subject in need of a therapeutic or prophylactic cancer treatment, wherein the paclitaxel is administered before or after administration to the subject To a method of treating healing or prophylactic cancer by administering to the subject a paclitaxel.

In one embodiment, the invention relates to a naphthofuran compound, or a pharmaceutically acceptable salt, solvate, hydrate or solvate thereof, which is referred to herein as "Compound 1 " Prophylactic or preventive cancer treatment in a human subject, which comprises administering a therapeutically effective amount of a prodrug,

(1)

(One)

In some embodiments, the compound is administered to a subject in a total daily dosage within the range of about 80 mg to about 2000 mg. In some embodiments, the compound is administered to a subject in a total daily dose selected from the group consisting of about 80 mg, about 160 mg, about 320 mg, about 480 mg, about 640 mg, about 800 mg and about 960 mg . In some embodiments, the compound is administered to the subject at a total daily dose of about 960 mg.

In some embodiments, the compound is administered twice daily (BID). In some embodiments, the compound is administered to a subject at a dose within the range of about 80 mg BID to about 480 mg BID. In some embodiments, the compound is administered to a subject in a dosage selected from the group consisting of about 80 mg BID, about 160 mg BID, about 320 mg BID, about 400 mg BID, and about 480 mg BID. In some embodiments, the compound is administered to a subject at a dose of about 480 mg BID.

In some embodiments, the compound is administered BID the time between administration of the compound is within the range of about 4 hours between administrations to about 16 hours between administrations. In some embodiments, the compound is administered to a subject in need thereof for a period of time greater than or equal to 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, and / or 16 hours. In some embodiments, the compound is administered to the subject at a dose within the range of about 80 mg BID to about 480 mg BID, with the time between administration of the compound being within the range of about 4 hours between administrations to about 16 hours between administrations. In some embodiments, the compound is administered to a subject in need thereof for a period of time greater than or equal to 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, and / or 16 hours. In some embodiments, the compound is administered to a subject in a dosage selected from the group consisting of about 80 mg BID, about 160 mg BID, about 320 mg BID, about 400 mg BID, and about 480 mg BID, The time is in the range of about 4 hours between administrations to about 16 hours between administrations. In some embodiments, the compound is administered to a subject in need thereof for a period of time greater than or equal to 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours, and / or 16 hours. In some embodiments, the compound is administered to a subject at a dose of about 480 mg BID, and the time between administration of the compound is about 12 hours between doses. In some embodiments, the compound is administered to a subject at a dose of about 80 mg BID, the time between administration of the compound being about 12 hours between doses. In some embodiments, the compound is administered to a subject at a dose of about 400 mg BID, and the time between administration of the compound is about 12 hours between doses. In some embodiments, the compound is administered to a subject at a dose of about 320 mg BID, and the time between administration of the compound is about 12 hours between doses. In some embodiments, the compound is administered to a patient at a time between about 4 hours and at least about 5 hours, at least about 6 hours, at least about 7 hours, at least about 8 hours, at least about 9 hours, at least about 10 hours, at least about 11 hours, Hour, at least about 12 hours, at least about 13 hours, at least about 14 hours, at least about 15 hours, and / or at least about 16 hours. In some embodiments, the compound is administered to a subject at a dose of about 80 mg BID, about 160 mg BID, about 320 mg BID, about 400 mg BID, and about 480 mg BID, the time between administration of the compound being at least 5 hours , Preferably between about 5 hours to about 15 hours between administrations.

In some embodiments, the compounds of the invention are administered as tablets or capsules. In some embodiments, the tablet or capsule contains a dose of about 80 mg.

In some embodiments, the compounds of the invention are orally administered with the fluids on an empty stomach. In some embodiments, the fluid is milk or water.

In some embodiments, the naphthofuran compound is a polymorph of the compound shown below referred to herein as "Compound 1"

(1)

(One)

For example, in some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-c] quinazoline which is characterized by an X-ray diffraction pattern substantially similar to that specified in WO 2011/116398 and WO 2011/116399. b] furan-4,9-dione, the contents of each of which are incorporated herein by reference in their entirety. The X-ray powder diffraction analysis shown in FIG. 1 of WO 2011/116398 and WO 2011/116399 showed Cu emission over a range of 5 ° to 70 ° with a step size of 0.03 ° at 40 KV / 30 mA and a count time of 3 hours Using a Philips PW1800 diffractometer used. The analysis was carried out from 2-45 ° 2θ using the following conditions: diverging slit: 0.6 mm, scattering slit: 0.6 mm, receiving slit: 0.1 mm, detector slit: 0.6 mm, step size: 0.02 °, 5 seconds. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-2-one, which is characterized by an X-ray diffraction pattern substantially similar to that specified in WO 2011/116398 and WO 2011/116399. 4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-2-one, which is characterized by an X-ray diffraction pattern substantially similar to that specified in WO 2011/116398 and WO 2011/116399. 4,9-dione. The X-ray powder diffraction analysis shown in Figures 2 and 3 of WO 2011/116398 and WO 2011/116399 was carried out using a Bruker D8 Advance diffractometer. The analysis was performed from 2-45 2 theta using the following conditions: diverging slit: 0.6 mm, scattering slit: 0.6 mm, receiving slit: 0.1 mm, detector slit: 0.6 mm, step size: 0.02, step time: 5 second.

For example, in some embodiments, the polymorphism is an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.4, 11.9, 14.1, 14.5, 17.3, 21.0, 22.2, 24.0, 26.0, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H, 9H, 9H, 9H, or 9H, characterized by an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.9, 14.1, 14.5, 17.3, -Naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 10.2 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 11.9 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.1 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 17.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 22.2 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.1 [ It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 10.2 degrees two theta, a peak at least about 11.9 degrees two theta, a peak at least about 14.1 degrees two theta, a peak at least about 14.5 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at at least about 22.2 DEG 2 & -Naphtho [2,3-b] furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X that comprises at least one peak at at least about 7.5, 9.9, 11.4, 12.3, 15.0, 23.0, 23.3, 24.1, 24.6, 25.0, 26.1, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, which is characterized by a line diffraction pattern. In some embodiments, the polymorphs are 2-acetyl-4H, which is characterized by an X-ray diffraction pattern comprising at least one peak at at least about 7.5, 9.9, 12.3, 15, 23.0, 23.3, 24.6 and / , 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 7.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 9.9 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 12.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 15 & It is a polymorph of Dion. In some embodiments, the polymorphism is a 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione which is characterized by an X-ray diffraction pattern comprising peaks at at least about 23 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 23.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 24.6 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.4 degrees two- It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 12.3 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at least about 23.3 DEG 2 &thetas; a peak at least about 24.6 DEG 2 DEG and at least about 28.4 DEG 2 theta and any combination thereof Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione.

The present invention also provides naphthofuran compounds in particulate form. For example, naphthofuran compounds in the form of particles are particles of a compound of formula I as shown below which are active, i. E., In vivo efficacy and / or antitumor activity. The effective particles or particles have a defined requirement for particle size and may have a defined requirement for particle size, for example, about 200, about 150, about 100, about 40 or about 20, about 10, about 5, About 3 占 퐉, about 2 占 퐉, about 1 占 퐉, about 0.5 占 퐉, or about 0.2 占 퐉 or less. Particles or particles larger than the defined particle size are inactive or less active.

In some embodiments, the compound comprises a population of compound particles, wherein a portion of the cumulative sum of the particles is present in the pharmaceutical composition having a diameter in the range of 0.2 [mu] m to 20 [mu] m.

In some embodiments, the compound comprises a population of compound particles, wherein 50% (D ₅₀ ) of the cumulative total of the particles is present in the pharmaceutical composition having a diameter within the range of about 0.5 to about 5 탆.

In some embodiments, the compound comprises a population of compound particles, wherein 50% (D ₅₀ ) of the cumulative total of the particles is present in the pharmaceutical composition having a diameter of about 2 μm.

In some embodiments, the naphthofuran compounds in particulate form are particles of a compound of formula (I) or a salt or solvate thereof, wherein the particles have a diameter of about 200 占 퐉 or less:

(I)

Wherein each R ₁ is independently selected from the group consisting of hydrogen, halogen, fluorine, cyano, nitro, CF ₃ , OCF ₃ , alkyl, methyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, Substituted aryl, OR &_lt; _{a >} , SR &_lt; _{a >} , and NH < _{2 >} , wherein R is selected from the group consisting of hydrogen, alkyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heterocycle, substituted heterocycle, ; n is 4; R ₃ is selected from the group consisting of hydrogen, halogen, fluorine, cyano, CF ₃ , OCF ₃ , alkyl, methyl, substituted alkyl, halogen-substituted alkyl, hydroxyl-substituted alkyl, Substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heterocycle, substituted heterocycle, aryl, substituted aryl, OR _a , SR _a, and NR _b R < _{c &} gt ;; R _a is selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heterocycle, Lt; / RTI > is independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, heterocycle, aryl and substituted aryl; R _b and R _c is independently selected from hydrogen, alkyl, the group consisting of substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocyclic, substituted heterocyclic, aryl, and substituted aryl, R _b and R _c is Together with the N to which they are attached form a heterocycle or a substituted heterocycle.

In some embodiments, each (R ₁ ) is independently selected from the group consisting of hydrogen, methyl, F (fluorine), Cl, Br, I, OH, and NH ₂ ; R ₃ is selected from the group consisting of methyl and C (R ₈ ) ₃ and each (R ₈ ) is independently selected from the group consisting of hydrogen, methyl, F (fluorine), Cl, Br, I, OH and NH ₂ Is selected. In some embodiments, no more than two of (R ₁ ) and (R ₈ ) are F (fluorine) and the remainder are hydrogen. In some embodiments, R < ₃ > is methyl. In a further embodiment, the compound is selected from the group consisting of 2- (1-hydroxyethyl) -naphtho [2,3-b] furan-4,9-dione, 2- acetyl- 2,3-b] furan-4,9-dione, 2-acetyl-7-fluoro-naphtho [ 4,9-dione, 2-ethyl-naphtho [2,3-b] furan-4,9-dione, its enantiomers, diastereoisomers, tautomers and salts or solvates.

In some embodiments, the naphthofuran compound in particle form is a particle of Compound 1.

In some embodiments, the naphthofuran compound in the form of particles is a polymorphic particle of compound 1. For example, in some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2 < RTI ID = 0.0 > , 3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] pyrimidine which is characterized by an X-ray diffraction pattern substantially similar to that specified in Figure 2 of WO 2011/116398 and WO 2011/116399 ] Furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H naphtho [2,3-b] thiophene which is characterized by an X-ray diffraction pattern substantially similar to that specified in Figure 3 of WO 2011/116398 and WO 2011/116399. Furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.4, 11.9, 14.1, 14.5, 17.3, 21.0, 22.2, 24.0, 26.0, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H, 9H, 9H, 9H, or 9H, characterized by an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.9, 14.1, 14.5, 17.3, -Naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs comprise 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 10.2 degrees two- . In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 11.9 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.1 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 17.3 DEG 2 & . In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 22.2 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.1 [ It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 10.2 degrees two theta, a peak at least about 11.9 degrees two theta, a peak at least about 14.1 degrees two theta, a peak at least about 14.5 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at at least about 22.2 DEG 2 & -Naphtho [2,3-b] furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X that comprises at least one peak at at least about 7.5, 9.9, 11.4, 12.3, 15.0, 23.0, 23.3, 24.1, 24.6, 25.0, 26.1, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, which is characterized by a line diffraction pattern. In some embodiments, the polymorphs are 2-acetyl-4H, which is characterized by an X-ray diffraction pattern comprising at least one peak at at least about 7.5, 9.9, 12.3, 15, 23.0, 23.3, 24.6 and / , 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 7.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 9.9 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 12.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 15 & It is a polymorph of Dion. In some embodiments, the polymorphism is a 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione which is characterized by an X-ray diffraction pattern comprising peaks at at least about 23 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 23.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 24.6 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.4 degrees two- It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 12.3 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at least about 23.3 DEG 2 &thetas; a peak at least about 24.6 DEG 2 DEG and at least about 28.4 DEG 2 theta and any combination thereof Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione.

In some embodiments, the particles have a diameter of about 160 microns, about 150 microns, about 120 microns, about 100 microns, about 50 microns, about 40 microns, or about 20 microns or less. In a further embodiment, the particles have a diameter of about 10, about 5, about 4, about 3, about 2, about 1, about 0.5, about 0.2 or about 0.1.

In some embodiments, the compound comprises a population of compound particles, wherein a portion of the cumulative sum of the particles is present in the pharmaceutical composition having a diameter in the range of 0.2 [mu] m to 20 [mu] m.

In some embodiments, the compound comprises a population of compound particles, wherein 50% (D ₅₀ ) of the cumulative total of the particles is present in the pharmaceutical composition having a diameter within the range of about 0.5 to about 5 탆.

In some embodiments, the compound comprises a population of compound particles, wherein 50% (D ₅₀ ) of the cumulative total of the particles is present in the pharmaceutical composition having a diameter of about 2 μm.

The present invention provides naphthofuran compounds, e. G. Particles or particles of a compound of formula (I) which are active, i. E. Have potency and / or antitumor activity. The active particles or particles have a specific size and may be of a size, for example, about 200 microns, about 150 microns, about 100 microns, about 40 microns, or about 20 microns, about 10 microns, about 5 microns, about 4 microns, about 3 microns, About 2 占 퐉, about 1 占 퐉, about 0.5 占 퐉, about 0.2 占 퐉, or about 0.1 占 퐉 or less. Particles or particles larger than a certain size are less active or inert than the particles described herein.

In some embodiments according to the present invention, the pharmaceutical composition comprises a compound, for example a naphthofuran of formula I or a salt or solvate thereof. For example, in some embodiments, the pharmaceutical composition comprises particles of Compound 1. For example, in some embodiments, the pharmaceutical composition comprises polymorphic particles of Compound 1. For example, in some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2 < RTI ID = 0.0 > , 3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] pyrimidine which is characterized by an X-ray diffraction pattern substantially similar to that specified in Figure 2 of WO 2011/116398 and WO 2011/116399 ] Furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b < RTI ID = 0.0 > ] Furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.4, 11.9, 14.1, 14.5, 17.3, 21.0, 22.2, 24.0, 26.0, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H, 9H, 9H, 9H, or 9H, characterized by an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.9, 14.1, 14.5, 17.3, -Naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 10.2 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 11.9 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.1 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 17.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 22.2 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.1 [ It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 10.2 degrees two theta, a peak at least about 11.9 degrees two theta, a peak at least about 14.1 degrees two theta, a peak at least about 14.5 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at at least about 22.2 DEG 2 & -Naphtho [2,3-b] furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X that comprises at least one peak at at least about 7.5, 9.9, 11.4, 12.3, 15.0, 23.0, 23.3, 24.1, 24.6, 25.0, 26.1, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, which is characterized by a line diffraction pattern. In some embodiments, the polymorphs are 2-acetyl-4H, which is characterized by an X-ray diffraction pattern comprising at least one peak at at least about 7.5, 9.9, 12.3, 15, 23.0, 23.3, 24.6 and / , 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 7.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 9.9 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 12.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 15 & It is a polymorph of Dion. In some embodiments, the polymorphism is a 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione which is characterized by an X-ray diffraction pattern comprising peaks at at least about 23 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 23.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 24.6 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.4 degrees two- It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 12.3 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at least about 23.3 DEG 2 &thetas; a peak at least about 24.6 DEG 2 DEG and at least about 28.4 DEG 2 theta and any combination thereof Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione.

A portion of the cumulative total of the particles may have a diameter of about 200 [mu] m or less. In some embodiments, a portion of the set of particles may be at least about 1%, at least about 5%, at least about 10%, at least about 20%, or at least about 30% of the total number of particles in the set. In some embodiments, the portion is a substantial fraction. For example, a "substantial portion" of a set of particles may comprise at least about 99%, at least about 95%, at least about 90%, at least about 85%, at least about 80%, at least about 75% Or more, about 60% or more, or about 50% or more. Each R ₁ is independently selected from the group consisting of hydrogen, halogen, fluorine, cyano, nitro, CF ₃ , OCF ₃ , alkyl, methyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl , Substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heterocycle, substituted heterocycle, aryl, substituted aryl, OR _a , SR _a and NH ₂ . n may be a positive integer; For example, n may be 4. R ₃ is selected from the group consisting of hydrogen, halogen, fluorine, cyano, CF ₃ , OCF ₃ , alkyl, methyl, substituted alkyl, halogen-substituted alkyl, hydroxyl-substituted alkyl, Substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heterocycle, substituted heterocycle, aryl, substituted aryl, OR _a , SR _a, and NR _b R < _c >. R _a is selected from the group consisting of hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heterocycle, Lt; / RTI > may be independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, heterocycle, aryl and substituted aryl. R _b and R _c may be independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, aryl and substituted aryl, or R _b and R _c together with the N to which they are attached form a heterocycle or a substituted heterocycle.

In some embodiments, according to the present invention, each (R ₁ ) may be independently selected from the group consisting of hydrogen, methyl, F (fluorine), Cl, Br, I, OH and NH ₂ . R ₃ may be selected from the group consisting of methyl and C (R ₈ ) ₃ . Each (R ₈ ) may be independently selected from the group consisting of hydrogen, methyl, F (fluorine), Cl, Br, I, OH and NH ₂ . In some embodiments, up to two of (R ₁ ) and R ₈ may be F (fluorine) and the remainder is hydrogen.

(I)

In some embodiments according to the present invention, the compound according to formula I is selected from the group consisting of 2- (1-hydroxyethyl) -naphtho [2,3-b] furan-4,9-dione, 2-acetyl- Naphtho [2,3-b] furan-4,9-dione, 2-acetyl-7-fluoro-naphtho [ , 3-b] furan-4,9-dione and 2-ethyl-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the compound according to Formula I is Compound 1. In some embodiments, the compound according to Formula I is a polymorph of Compound 1. For example, in some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2 < RTI ID = 0.0 > , 3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] pyrimidine which is characterized by an X-ray diffraction pattern substantially similar to that specified in Figure 2 of WO 2011/116398 and WO 2011/116399 ] Furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b < RTI ID = 0.0 > ] Furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.4, 11.9, 14.1, 14.5, 17.3, 21.0, 22.2, 24.0, 26.0, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H, 9H, 9H, 9H, or 9H, characterized by an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.9, 14.1, 14.5, 17.3, -Naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 10.2 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 11.9 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.1 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 17.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 22.2 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.1 [ It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 10.2 degrees two theta, a peak at least about 11.9 degrees two theta, a peak at least about 14.1 degrees two theta, a peak at least about 14.5 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at at least about 22.2 DEG 2 & -Naphtho [2,3-b] furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X that comprises at least one peak at at least about 7.5, 9.9, 11.4, 12.3, 15.0, 23.0, 23.3, 24.1, 24.6, 25.0, 26.1, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, which is characterized by a line diffraction pattern. In some embodiments, the polymorphs are 2-acetyl-4H, which is characterized by an X-ray diffraction pattern comprising at least one peak at at least about 7.5, 9.9, 12.3, 15, 23.0, 23.3, 24.6 and / , 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 7.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 9.9 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 12.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 15 [ . In some embodiments, the polymorphism is a 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione which is characterized by an X-ray diffraction pattern comprising peaks at at least about 23 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 23.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 24.6 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.4 degrees two- It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 12.3 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at least about 23.3 DEG 2 &thetas; a peak at least about 24.6 DEG 2 DEG and at least about 28.4 DEG 2 theta and any combination thereof Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione.

For example, the pharmaceutical composition may have a particle size of at least about 90% of the cumulative total of particles of about 160, 100, 40, 20, 10, 5, 3 or 2 microns . For example, the pharmaceutical composition may have a particle size distribution wherein about 50% or more of the cumulative total of the particles is about 160, 100, 40, 20, 10, 5, 3, 2, Particle size. For example, the pharmaceutical composition may have a particle size of at least about 10% of the cumulative total of the particles of about 160, 100, 40, 20, 5, 2, 1, 0.5, Lt; / RTI > In the pharmaceutical composition, the particles have a median diameter of, for example, about 160 탆, 40 탆, 20 탆, 10 탆, 5 탆, 4 탆 3 탆, 2 탆, 1 탆, 0.5 탆, 0.3 탆, Lt; / RTI > For example, the particles may have a median diameter of about 0.2 microns to about 50 microns, or a median diameter of about 0.5 microns to about 30 microns. For example, the pharmaceutical composition has a cumulative sum of particles having a ratio of mean diameter to median diameter of about 2 占 퐉 or less. The pharmaceutical composition may be in a crystalline state and may have particles comprising the compound in at least two different polymorphic states.

In some embodiments, the pharmaceutical composition comprises a compound of Formula I, or a polymorph thereof, in particulate form, wherein the particles or particles are 20 microns, 10 microns, 5 microns, 2 microns, 1 micron, or less than 0.5 microns.

The present invention provides substantially pure compounds of formula II:

(II)

(II)

In the above formulas, each R < ₁ > is independently H, Cl or F; n is 0, 1, 2, 3 or 4; In some embodiments, the compound of formula II is present in particulate form.

In some embodiments, the substantially pure compound is Compound 1. In some embodiments, Compound 1 is present in particulate form.

In some embodiments, the substantially pure compound is 2- (1-hydroxyethyl) -naphtho [2,3-b] furan-4,9-dione, 2- acetyl-7- chloronaphtho [2,3 -b] furan-4,9-dione, 2-acetyl-7-fluoro-naphtho [2,3- b] furan-4,9-dione, 2- acetylnaphtho [2,3- b] furan 4,9-dione, 2-ethyl-naphtho [2,3-b] furan-4,9-dione, mono - [1- (4,9-dioxo-3a, 2-yl) -vinyl] ester, phosphoric acid 1- (4,9-dioxo-3a, 4,9,9a-tetrahydro-naphtho [2, 3-b] furan-2-yl) -vinyl ester dimethyl ester, its enantiomers, diastereoisomers, tautomers and salts or solvates thereof.

In some embodiments, the substantially pure compound is a polymorph of compound 1. [ For example, in some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2 < RTI ID = 0.0 > , 3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] pyrimidine which is characterized by an X-ray diffraction pattern substantially similar to that specified in Figure 2 of WO 2011/116398 and WO 2011/116399 ] Furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b < RTI ID = 0.0 > ] Furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.4, 11.9, 14.1, 14.5, 17.3, 21.0, 22.2, 24.0, 26.0, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H, 9H, 9H, 9H, or 9H, characterized by an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.9, 14.1, 14.5, 17.3, -Naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs comprise 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 10.2 degrees two- . In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 11.9 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.1 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 17.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 22.2 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.1 [ It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 10.2 degrees two theta, a peak at least about 11.9 degrees two theta, a peak at least about 14.1 degrees two theta, a peak at least about 14.5 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at at least about 22.2 DEG 2 & -Naphtho [2,3-b] furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X that comprises at least one peak at at least about 7.5, 9.9, 11.4, 12.3, 15.0, 23.0, 23.3, 24.1, 24.6, 25.0, 26.1, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, which is characterized by a line diffraction pattern. In some embodiments, the polymorphs are 2-acetyl-4H, which is characterized by an X-ray diffraction pattern comprising at least one peak at at least about 7.5, 9.9, 12.3, 15, 23.0, 23.3, 24.6 and / , 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 7.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 9.9 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 12.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 15 & It is a polymorph of Dion. In some embodiments, the polymorphism is a 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione which is characterized by an X-ray diffraction pattern comprising peaks at at least about 23 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 23.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 24.6 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.4 degrees two- It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 12.3 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at least about 23.3 DEG 2 &thetas; a peak at least about 24.6 DEG 2 DEG and at least about 28.4 DEG 2 theta and any combination thereof Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione.

In some embodiments, the polymorph of Compound 1 is present in particulate form.

In some embodiments, the compound, product and / or pharmaceutical composition has a purity of at least about 80%, about 85%, about 90%, about 95%, or about 99%. In some embodiments, the compound, product and / or pharmaceutical composition comprises at least about 95.5%, about 96%, about 96.5%, about 97%, about 97.5%, about 98%, about 98.5%, about 99% %. ≪ / RTI > In some embodiments, the compound, product and / or pharmaceutical composition comprises at least about 99.1%, about 99.2%, about 99.3%, about 99.4%, about 99.5%, about 99.6%, about 99.7%, about 99.8% %. ≪ / RTI >

In some embodiments, the compound, product, and / or pharmaceutical composition has less than about 10%, about 5%, about 1%, about 0.15%, or about 0.15% impurity. In some embodiments, the compound, product, and / or pharmaceutical composition has less than about 0.5%, about 0.2%, about 0.15%, or about 0.1% impurity. In a further embodiment, the impurity is selected from the group consisting of 2-acetyl-2,3-dihydronaphtho [2,3-b] furan-4,9-dione, 2,6-diacetylnaphtho [ 2,3-b] furan-4,9-dione, 2,7-diacetyl-naphtho [2,3- 2,3-b] furan-4,9-dione, naphtho [2,3-b] furan- Diol and 1- (4,9-dihydroxy-naphtho [2,3-b] furan-2-yl) -ethanone.

In some embodiments, the impurity comprises a residual solvent. In some embodiments, the solvent is selected from the group consisting of ethyl acetate (EtOAc), toluene, ethanol, methanol, chloroform and CH ₂ Cl ₂ / hexane.

In some embodiments, the purity is measured by HPLC (high performance liquid chromatography). In some embodiments, the purity is measured by NMR (nuclear magnetic resonance). In a further embodiment, the purity is measured in both HPLC and NMR.

The present invention also provides a polymorph of Compound 1 in particulate form, wherein the compound is a highly purified form of the product and / or pharmaceutical composition. For example, in some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2 < RTI ID = 0.0 > , 3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] pyrimidine which is characterized by an X-ray diffraction pattern substantially similar to that specified in Figure 2 of WO 2011/116398 and WO 2011/116399 ] Furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b < RTI ID = 0.0 > ] Furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.4, 11.9, 14.1, 14.5, 17.3, 21.0, 22.2, 24.0, 26.0, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H, 9H, 9H, 9H, or 9H, characterized by an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.9, 14.1, 14.5, 17.3, -Naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 10.2 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 11.9 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.1 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 17.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 22.2 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.1 [ It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 10.2 degrees two theta, a peak at least about 11.9 degrees two theta, a peak at least about 14.1 degrees two theta, a peak at least about 14.5 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at at least about 22.2 DEG 2 & -Naphtho [2,3-b] furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X that comprises at least one peak at at least about 7.5, 9.9, 11.4, 12.3, 15.0, 23.0, 23.3, 24.1, 24.6, 25.0, 26.1, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, which is characterized by a line diffraction pattern. In some embodiments, the polymorphs are 2-acetyl-4H, which is characterized by an X-ray diffraction pattern comprising at least one peak at at least about 7.5, 9.9, 12.3, 15, 23.0, 23.3, 24.6 and / , 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 7.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 9.9 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 12.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 15 & It is a polymorph of Dion. In some embodiments, the polymorphism is a 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione which is characterized by an X-ray diffraction pattern comprising peaks at at least about 23 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 23.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 24.6 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.4 degrees two- It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 12.3 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at least about 23.3 DEG 2 &thetas; a peak at least about 24.6 DEG 2 DEG and at least about 28.4 DEG 2 theta and any combination thereof Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione.

The polymorph of Compound 1 exists in the form of particles. In some embodiments, the polymorph of Compound 1 is present in particulate form and the particles have a diameter of about 160 microns, about 150 microns, about 120 microns, about 100 microns, about 50 microns, about 40 microns, or about 20 microns . In some embodiments, the polymorph of Compound 1 in particle form is present in a population of particles, and the population of particles is about 160 [mu] m, about 150 [mu] m, about 120, about 100, about 50, about 40 [ 20 ㎛ D ₅₀ or less has the (i.e., the center point of the second distribution of the particle size distribution is split into equal parts). In some embodiments, the polymorph of Compound 1 is present in the form of particles, and the particles can be about 10 탆, about 5 탆, about 4 탆, about 3 탆, about 2 탆, about 1 탆, about 0.5 탆, Or a diameter of about 0.1 mu m or less. In some embodiments, the polymorph of Compound 1 in particle form is in a population of particles, and the population of particles is about 10, about 5, about 4, about 3, about 2, about 1, It has a 0.5 ㎛ or D ₅₀ of less than or equal to about 0.2 ㎛.

The present invention provides a population of particles or particles of compound 1 having activity, i. E., Potency and / or antitumor activity. The active particles or particles may be of a particular size, for example, about 200, about 150, about 100, about 40 or about 20, about 10, about 5, and it has a diameter of D ₅₀ or greater than about 1 ㎛, about 0.5 or about 0.2 ㎛ ㎛. Particles or particles larger than a certain size are less active or inert than the particles described herein.

A portion of the cumulative total of the polymorphic particles of Compound 1 may have a diameter or D ₅₀ of about 200 μm or less. In some embodiments, a portion of the set of particles may be at least about 1%, at least about 5%, at least about 10%, at least about 20%, or at least about 30% of the total number of particles in the set. In some embodiments, the portion is substantial. For example, a "substantial portion" of a set of particles may comprise at least about 99%, at least about 95%, at least about 90%, at least about 85%, at least about 80%, at least about 75% , Greater than 70%, greater than about 60%, or greater than about 50%.

In some embodiments, the population of polymorphic particles of Compound 1 is such that at least about 90% of the cumulative total of the particles is about 160, 100, 40, 20, 10, 5, 3, 1 < / RTI > or 0.5 mu m or less. For example, a population of polymorphic particles of Compound 1 may have a particle size distribution such that at least about 50% of the cumulative total of particles is greater than about 160, 100, 40, 20, 10, 5, 3, Mu m or 0.5 mu m or less. For example, a population of polymorphic particles of Compound 1 may have a particle size distribution such that at least about 10% of the cumulative total of particles is greater than about 160, 100, 40, 20, 5, 2, Lt; RTI ID = 0.0 > um. ≪ / RTI > In the population of polymorphic particles of Compound 1, the particles can be, for example, about 160, 40, 20, 10, 5, 4, 3, 2, 1, Lt; RTI ID = 0.0 > diameter. For example, the particles may have a median diameter of about 0.002 μm to about 50 μm, or a median diameter of about 0.2 μm to about 30 μm. For example, a population of polymorphic particles of Compound 1 may have a cumulative sum of particles having a ratio of mean diameter to median diameter of about 2 or less. The population of polymorphic particles of Compound 1 can be in a crystalline state and can have particles comprising compounds in two different polymorphic states.

In some embodiments, the polymorph of Compound 1 is present in particulate form and the particles have a diameter of about 20 microns, 10 microns, 5 microns or 23 microns, 2 microns, 1 micron, 0.5 microns, 0.2 microns, or 0.1 microns or less Respectively. In some embodiments, the polymorph of Compound 1 in particulate form is present in a population of particles, wherein the population of particles is about 20 microns, 10 microns, 5 microns, 4 microns, 5 microns, 3 microns, 2 microns, has a 0.5 micron or a D ₅₀ of less than 0.2 micron.

The present invention also provides a pharmaceutical composition comprising a therapeutically effective amount of a substantially pure naphthofuran compound and a pharmaceutically acceptable carrier, excipient or diluent. Examples of excipients include glycerol esters of fatty acids, glycerol esters of saturated fatty acids, glycerol esters of saturated fatty acids having 8 to 18 carbons, glyceryl laurate, polyethylene glycol, cellulose, microcrystalline cellulose, carboxymethyl cellulose, phosphatidylcholine, Lipids, sterols, cholesterol, surfactants, polysorbates and / or polyoxyethylene sorbitan alkylates.

In some embodiments, according to the present invention, the article of manufacture may comprise a container containing a therapeutically effective amount of the pharmaceutical composition and a pharmaceutically acceptable excipient.

Methods of making compounds, products and / or pharmaceutical compositions according to some embodiments of the present invention may include milling the compounds to form particles. For example, the compound may be processed by ball milling, roll milling, jet milling, wet milling, ultrasonic milling, milling or any combination thereof and / or other milling procedures. The temperature of the compound can be reduced, for example reduced to a cryogenic temperature, and can be milled. This reduction in temperature allows the compound to break better and allows for a better reduction of particle size by milling.

Methods of making compounds, products and / or pharmaceutical compositions according to some embodiments of the present invention may include crystallization. The particle size distribution (PSD) obtained during crystallization is affected by the combination of various mechanisms that occur during crystallization, such as nucleation, growth, cohesion, wear, fracture, and the like. If the particle size can not be constantly controlled during crystallization to meet desired specifications, it may include additional processing steps, such as dry milling.

In some embodiments of the invention, a composition for reducing or inhibiting the replication or spread of neoplastic cells comprises a set of particles selected by the following method. The compounds according to formula I or salts or solvates thereof may be provided:

(I)

In some embodiments, Compound 1 or a salt or solvate thereof may be provided. In some embodiments, the polymorph of compound 1 may be provided. For example, in some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2 < RTI ID = 0.0 > , 3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] pyrimidine which is characterized by an X-ray diffraction pattern substantially similar to that specified in Figure 2 of WO 2011/116398 and WO 2011/116399 ] Furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b < RTI ID = 0.0 > ] Furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.4, 11.9, 14.1, 14.5, 17.3, 21.0, 22.2, 24.0, 26.0, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H, 9H, 9H, 9H, or 9H, characterized by an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.9, 14.1, 14.5, 17.3, -Naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 10.2 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 11.9 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.1 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 17.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 22.2 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.1 [ It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 10.2 degrees two theta, a peak at least about 11.9 degrees two theta, a peak at least about 14.1 degrees two theta, a peak at least about 14.5 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at at least about 22.2 DEG 2 & -Naphtho [2,3-b] furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X that comprises at least one peak at at least about 7.5, 9.9, 11.4, 12.3, 15.0, 23.0, 23.3, 24.1, 24.6, 25.0, 26.1, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, which is characterized by a line diffraction pattern. In some embodiments, the polymorphs are 2-acetyl-4H, which is characterized by an X-ray diffraction pattern comprising at least one peak at at least about 7.5, 9.9, 12.3, 15, 23.0, 23.3, 24.6 and / , 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 7.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 9.9 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 12.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 15 & It is a polymorph of Dion. In some embodiments, the polymorphism is a 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione which is characterized by an X-ray diffraction pattern comprising peaks at at least about 23 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 23.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 24.6 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.4 degrees two- It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 12.3 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at least about 23.3 DEG 2 &thetas; a peak at least about 24.6 DEG 2 DEG and at least about 28.4 DEG 2 theta and any combination thereof Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione.

One or more sets of particles comprising the compound. The particle size distribution of each of at least one set of particles can be measured. One or more sets of particles may be administered to the neoplastic and normal cells for a predetermined time at a predetermined concentration. The effect of particles on metabolism and / or division of neoplastic and normal cells can be observed. Efficiency assessments can be assigned to each set of particles based on the effect of the particles on the neoplastic cell. The toxicity assessment can be assigned to each set of particles based on the effect of the particles on normal cells. An efficiency assessment and / or a toxicity assessment of one or more sets of particles having a first particle size distribution may be made by evaluating the efficiency and / or toxicity of one or more other sets of particles having a particle size distribution different from the first particle size distribution ≪ / RTI > A set of particles with a greater efficiency rating than the one or more other sets of particles, less toxicity assessment, and / or a larger weighted efficacy assessment and toxicity evaluation sum may be selected as the optimal set. For example, the particle size distribution of the optimal set of particles can be identified as the optimal particle size distribution. For example, an optimal set of particles may be included in the composition. For example, an efficiency assessment may be proportional to the antitumor activity. For example, an efficiency assessment may be based on inhibition of metabolism and / or division of neoplastic cells. For example, toxicity assessment may be inversely proportional to tolerability. For example, toxicity assessment may be based on inhibition of metabolism and / or division of normal cells. For example, one or more sets of particles may be administered in vitro to neoplastic and normal cells. For example, an efficiency assessment may be based on the IC ₅₀ of neoplastic cells. For example, toxicity assays can be based on the IC ₅₀ of normal cells. For example, one or more sets of particles can be administered in vivo to neoplastic cells and normal cells in a test animal. Test animals can be, for example, mammals, primates, rats, rats, guinea pigs, rabbits or dogs. The efficiency assessment may be based on a reduction in the volume of neoplastic cells and the toxicity assessment may be based on a reduction in the mass of the test animal.

In some embodiments, the preparation of a set of particles comprising a compound includes dissolving and dispersing the compound, dissolving and dispersing the compound using microfluidic techniques, dissolving and dispersing the compound using cavitation or spraying, compound milling, compound , Ball milling of the compound, roll milling of the compound, jet milling of the compound, wet milling of the compound, ultrasonic milling of the compound, milling of the compound and / or sieving of the compound. The particles may be suspended in a pharmaceutically acceptable excipient. The determination of the particle size distribution may include using techniques selected from the group consisting of sieve analysis, optical microscopy, electron microscopy, electrical resistivity, settling time, laser diffraction, acoustic spectroscopy, and combinations .

The method of treating neoplasia or other cell proliferative disorders comprises administering a therapeutically effective amount of a composition comprising an optimal set of particles of a composition having an optimal particle size and distribution to a person suffering from a neoplasm, a mammal or an animal .

The present invention provides a process for preparing a naphthofuran compound. The process comprises reacting a mixture comprising a naphthodihydrofuran compound or a naphthodihydrofuran compound with an oxidizing agent in a first solvent. In some embodiments, the mixture further comprises a naphthofuran compound. In some embodiments, the naphthofuran compound is selected from the group consisting of 2- (1-hydroxyethyl) -naphtho [2,3-b] furan-4,9-dione, 2- acetyl- 2,3-b] furan-4,9-dione, 2-acetylnaphtho [2,3-b] Furan-4,9-dione, 2-ethyl-naphtho [2,3-b] furan-4,9-dione, mono - [1- (4,9-dioxo-3a, 2-yl) -vinyl] ester, phosphoric acid 1- (4,9-dioxo 36 3a, 4,9,9a-tetrahydro-naphtho [2 , 3-b] furan-2-yl) -vinyl ester dimethyl ester, its enantiomers, diastereoisomers, tautomers and salts or solvates thereof. In some embodiments, the oxidizing agent is manganese dioxide. In some embodiments, the first solvent is toluene. In some embodiments, the method further comprises filtering the oxidation product onto a pad of activated carbon. In some embodiments, the method further comprises evaporating the first solvent to crystallize the naphthofuran compound. In some embodiments, the method further comprises recrystallizing the naphthofuran compound using a second solvent. In some embodiments, the second solvent is ethyl acetate. In some embodiments, the method further comprises slurrying the naphthofuran compound using a second solvent, heating the slurry, and cooling the slurry.

The present invention provides a process for preparing a substantially pure naphthofuran compound. The process comprises crystallizing a naphthofuran compound using a first solvent and recrystallizing the naphthofuran compound using a second solvent. The present invention provides another process for preparing a substantially pure naphthofuran compound. The method includes crystallizing the naphthofuran compound using a first solvent, making the crystalline naphthofuran compound a slurry using a second solvent, heating the slurry, and cooling the slurry. In some embodiments, the naphthofuran compound is 2- (1-hydroxyethyl) -naphtho [2,3-b] furan-4,9-dione, 2- acetyl-7- chloronaphtho [2,3 -b] furan-4,9-dione, 2-acetyl-7-fluoro-naphtho [2,3- b] furan-4,9-dione, 2- acetylnaphtho [2,3- b] furan 4,9-dione, 2-ethyl-naphtho [2,3-b] furan-4,9-dione, mono - [1- (4,9-dioxo-3a, 2-yl) -vinyl] ester, phosphoric acid 1- (4,9-dioxo-3a, 4,9,9a-tetrahydro-naphtho [2, 3-b] furan-2-yl) -vinyl ester dimethyl ester, its enantiomers, diastereoisomers, tautomers and salts or solvates thereof. In some embodiments, the first solvent is toluene. In some embodiments, the second solvent is ethyl acetate.

The present invention provides naphthofuran compounds produced by any one of the above methods. In some embodiments, the naphthofuran compound is selected from the group consisting of 2- (1-hydroxyethyl) -naphtho [2,3-b] furan-4,9-dione, 2- acetyl- 2,3-b] furan-4,9-dione, 2-acetylnaphtho [2,3-b] Furan-4,9-dione, 2-ethyl-naphtho [2,3-b] furan-4,9-dione, mono - [1- (4,9-dioxo-3a, 2-yl) -vinyl] ester, phosphoric acid 1- (4,9-dioxo-3a, 4,9,9a-tetrahydro-naphtho [2 , 3-b] furan-2-yl) -vinyl ester dimethyl ester, its enantiomers, diastereoisomers, tautomers and salts or solvates thereof. In some embodiments, the naphthofuran compound has a purity of at least about 80%, about 85%, or about 90%, about 95%, or about 99%. In some embodiments, the naphthofuran compound has a purity of about 10%, about 5%, about 2%, or about 1%, about 0.5%, about 0.2%, about 0.15%, or about 0.1% or less.

The present invention provides a method for preparing particles of Compound 1 comprising polymorphic particles of Compound 1, particles of very pure form of Compound 1, and polymorphically very pure forms of Compound 1. In some embodiments, the particles having a predetermined median particle size, for example, about 20 microns, are produced by milling the crystals of Compound 1 and mixing the crystals of the purified form of Compound 1, the polymorphic crystals of Compound 1 and / ≪ / RTI > of the crystals of Compound 1, which comprises crystals of the polymorph in the form of crystals. For example, the crystals are milled using a jet milling method with a venturi pressure of about 40, a milling pressure of about 100, and a feed rate of about 1,304 g / hr.

The present invention also provides kits and / or methods for the treatment of a particular selected patient population suitable for therapeutic administration of a compound of this disclosure by detecting the level of expression of one or more biomarkers associated with cancer stem cells. The biomarker can be compared with levels in patients not suffering from cancer known to have baseline, control or normal levels of expression of the same marker, such as known to have cancer stem cells and / or more Stat3 pathway activity , Cancer stem cells that are known to have cancer stem cells and / or whose expression is increased in patients or patients suffering from cancer known to have stat3 pathway activity.

In some embodiments, the biomarker associated with cancer stem cells is phosphorylated STAT3 (p-STAT3). In some embodiments, the biomarker associated with cancer stem cells is beta -catenin. In some embodiments, the biomarker associated with cancer stem cells is NANOG. In some embodiments, a combination of biomarkers associated with cancer stem cells is used, and the combination is selected from the group consisting of two or more of p-STAT3, beta -catenin, and nanox. In some embodiments, a combination of biomarkers associated with cancer stem cells is used, and the combination is selected from the group consisting of three species of p-STAT3, beta -catenin, and nanorh.

In the methods and / or kits of this disclosure, the level of expression of one or more cancer stem cell markers is detected in a patient or in a sample from a patient, wherein the patient or sample has one or more cancer stem cell markers , So that the patient administers a therapeutically effective amount of a compound of this disclosure.

In some embodiments of these methods, the method is an in vivo method. In some embodiments of these methods, the method is an in situ method. In some embodiments of these methods, the method is an in vitro method. In some embodiments of these methods, the method is an in vitro method.

The invention also provides a kit for identifying, or otherwise purifying, e.g., stratifying, a population of patients suitable for therapeutic administration of a compound of this disclosure by detecting the level of expression of one or more biomarkers associated with cancer stem cells and / ≪ / RTI > The biomarker can be compared with levels in patients not suffering from cancer known to have baseline, control or normal levels of expression of the same marker, such as known to have cancer stem cells and / or more Stat3 pathway activity , Cancer stem cells that are known to have cancer stem cells and / or whose expression is increased in patients or patients suffering from cancer known to have stat3 pathway activity.

In some embodiments, the biomarker associated with cancer stem cells is phosphorylated STAT3 (p-STAT3). In some embodiments, the biomarker associated with cancer stem cells is beta -catenin. In some embodiments, the biomarker associated with cancer stem cells is lanthanum. In some embodiments, a combination of biomarkers associated with cancer stem cells is used and the combination is selected from the group consisting of at least two of p-STAT3, beta -catenin, and nanox. In some embodiments, a combination of biomarkers associated with cancer stem cells is used, the combination being p-STAT3, beta -catenin, and nanoporus.

In the methods and / or kits of the present disclosure, the level of expression of one or more cancer stem cell markers is detected in a patient or in a sample from a patient, wherein the patient or sample has one or more cancer stem cell markers And the patient administers a therapeutically effective amount of a compound of the present disclosure.

In some embodiments of these methods, the method is an in vivo method. In some embodiments of these methods, the method is an in situ method. In some embodiments of these methods, the method is an in vitro method. In some embodiments of these methods, the method is an in vitro method.

Figure 1 is a graph comparing the pharmacokinetics of BID administration versus QD administration in a patient, with 500 mg administered to a patient during each administration. Doses were administered at 4 hour intervals between two doses in the same primary for 500 mg BID therapy.
Figure 2 is a graph comparing the pharmacokinetics of two different agents of the compounds of the present invention. The two formulations produce capsules of different sizes.
Figure 3a consists of photographic images of tumor tissue samples from CRC patients visualized by immunohistochemistry using antibodies against phosphorylated STAT3 and DAPI (bottom line).
Figure 3b is a chart showing trends for improvement in survival for patients with high p-STAT3 (versus patients with low or negative p-STAT3).
4A is a photomicrograph of tumor tissue samples from CRC patients visualized by immunohistochemistry using antibodies against [beta] -catenin and DAPI (dorsal raphe).
Figure 4B is a chart showing trends for improvement in survival for patients with nuclear beta -catenin localization (compared to patients with cell membrane-specific beta -catenin).
Figure 5 shows CD44 ^high cell growth blocked by compounds of the invention. CD44 ^solid cells were isolated by FACS (FaDu) and were incubated for 5 days in the absence of adherence and serum to form primary sphere. Thereafter, the primary sphere was digested into single cells in Accumax (eBioscience, San Diego, CA), cultured for 72 hours as above, and then the indicated concentration of therapeutic agent was added . Five days after the treatment, representative representative images were taken.
Figure 6 shows an in vivo experiment of nude mice with xenografted human colorectal tumor tissue showing that the compounds of the present invention are effective in reducing or clearing p-STAT3 and beta -catenin levels. Formaldehyde-fixed tumors from mice treated daily for 15 days with oral feeding of compounds of the present invention or vehicle (control) were cut and fixed in immunofluorescent staining using antibodies specific for human STAT2 and < RTI ID = 0.0 > Respectively.
FIG. 7 shows that the compound of the present invention targeted cancer stem cells in a mouse experiment. Mice with xenotransplantation were intraperitoneally administered the vehicle of the present invention with vehicle, gemcitabine (120 mg / kg (MIA PaCa-2)), carboplatin (30 mg / kg (FaDu)) or 20 mg / kg . Mice were sacrificed, and tumors were collected 7 or 14 days after treatment on PaCa-2 and FaDu cells, respectively. After animal sacrifice and sterilization of the tumor, a single cell suspension was obtained. Thereafter, live cells were counted and cultured in cancer stem cell medium (DMEM / F12, B27 Neurobasal supplement, 20 ng / ml EGF, 10 ng / ml FGF, 4 ng / ml insulin and 0.4% BSA) Was used to measure his ability to form spheres. New medium was added every 3 days and spheroid formation was measured after 10-14 days in culture. ≫ 50 cells were recorded.
Figures 8a, 8b and 8c are a series of graphs showing that in human clinical trials the compounds of the present invention were found to be effective in CRC patients. Figure 8a shows the relationship between progression-free survival (PFS) and exposure of compounds of the invention in patients with colorectal cancer (CRC). In CRC patients, statistical significance differences appeared in PFS between patients with compound plasma concentrations of the invention of greater than 2.0 [mu] M over 4 hours and patients not reaching the level of exposure. Figure 8b shows overall survival (OS) in assessable CRC patients. Past controls [Cetuximab for the treatment of colorectal cancer, 2007, N. Engl . J. Med . OS of an evaluable CRC patient treated with a compound of the present invention (defined as a compound of the present invention > = 4 weeks, 80% satisfied) compared to the control group. Figure 8C shows the PFS in an evaluable CRC patient. Past controls [Chemotherapy - open-label Phase III trials of panni tomus plus optimal support therapy compared with optimal adjuvant therapy alone in patients with refractory metastatic colorectal cancer, 2007, J. Clin . Onc . PFS of an evaluable CRC patient treated with a compound of the present invention (defined as a compound of the present invention > = 4, 80% satisfied) compared to the PFS of the present invention (25: 1658-1665).

Embodiments of the present invention are discussed in detail below. In the description of the embodiments, certain terminology is used for the sake of clarity. However, the present invention is not intended to be limited to the specific terms so selected. Those skilled in the relevant art will recognize that other equivalent components may be used and other methods may be developed without departing from the spirit and scope of the invention. All references cited herein are incorporated by reference as if each individual publication were individually incorporated.

As used herein, a "substantial fraction" of a set of particles is at least about 99%, at least about 95%, at least about 90%, at least about 85%, at least about 80% , At least about 70%, at least about 60%, or at least about 50%.

The anticancer stem cell activity of the composition can be measured in vitro or in vivo. For example, the antitumor activity of the composition can be measured in vitro by administering the compound and measuring the self-renewal and survival of cancer stem cells. For example, the antitumor activity of a compound can be evaluated in vitro by comparing the pattern of the tumor cells to which the compound is administered with the pattern of the tumor cells (control) without administration of the compound. For example, the antitumor activity of a composition can be measured in vivo by measuring the volume change of the tumor in the animal to which the compound is administered, applying a metastatic model, and / or applying an isothermal model. For example, the antitumor activity of a compound can be assessed in vivo by comparing an animal to which the compound is administered and an animal to which no compound is administered (control).

The tolerability of the composition can be determined in vitro or in vivo. For example, the compound may be administered, the rate of cleavage of normal cells may be measured, the nutrient uptake of normal cells may be measured, the index of normalized metabolic rate of normal cells may be measured, Or another index of vitality of normal cells may be measured to determine the tolerability of the composition in vitro. For example, the tolerability of a compound can be assessed in vitro by comparing aspects of normal cells to which the compound has been administered and aspects of normal cells (control) without administration of the compound. For example, the tolerability of the composition can be determined in vivo by measuring body weight or food intake in an animal to which the compound is administered, or by conducting clinical observations such as reactivity to hair retention or loss, activity and / or stimulation. For example, the tolerability of a compound can be assessed in vivo by comparing an animal that received the compound with an animal that did not receive the compound (control).

The compounds, products and / or pharmaceutical compositions may be assigned an efficiency and / or toxicity assessment. For example, an efficacy assessment may be proportional to anti-tumor activity or may be a monotone increasing function with respect to anti-tumor activity. For example, a toxicity assessment may be inversely related to tolerability or may be a monotonically decreasing function with respect to tolerability. It has been reported that the naphthofuran compounds are deficient in in vivo antitumor activity. See MM Rao and DGI Kingston, J. Natural Products , 45 (5) (1982) 600-604. Furthermore, compounds have been reported to have equivalent toxicity to cancer cells and normal cells. That is, the compounds have been reported to kill cancer cells and normal cells equally, which can lead to the conclusion that the compounds are unlikely to be able to treat cancer. See K. Hirai K. et al., Cancer Detection and Prevention , 23 (6) (1999) 539-550; Takano A. et al., Anti-cancer Research 29: 455-464, 2009].

However, the experimental studies reported herein demonstrate that compounds have selective antineoplastic activity upon administration of the compound as particles having an appropriate particle size distribution to achieve a particular pharmacokinetic exposure, as described in this document.

For purposes of the present invention, the "bioavailability" of a drug is defined as the relative amount of drug from the administered dosage form going to the systemic cycle and the rate at which the drug appears in the blood stream. Bioavailability depends on three or more factors: (i) absorption to regulate bioavailability, (ii) tissue re-distribution and (iii) elimination (metabolic degradation and elongation and other mechanisms).

"Absolute bioavailability" is estimated with reference to tissue redistribution and bioconversion (ie, elimination) that can be estimated by intravenous administration of the drug. Unless otherwise indicated, "HPLC" refers to high performance liquid chromatography; "Pharmaceutically acceptable" refers to a physiologically acceptable substance that upon administration to a mammal does not normally produce an allergic or other adverse reaction, such as an acute gastrointestinal disorder, dizziness, or the like; "Mammal" refers to a class of higher vertebrates, including mammals secreted by mammary glands to their offspring, and generally those whose skin is somewhat covered with hair and all other animals; "Treating" is intended to include relaxing, alleviating or eliminating one or more symptoms of the disease (s) in the mammal.

The term "treating" as used herein is intended to include the administration of a compound according to the invention to prophylactically prevent or inhibit undesired conditions and therapeutically eliminate or reduce the degree or symptomatology of the condition. Treatment also includes preventing recurrence of the unwanted condition, delaying the progression of the unwanted condition, and preventing or delaying the onset of the unwanted condition. The treatment according to the invention provides a person or other mammal with the disease or condition that produces the need for treatment. Treatment also includes in vitro application of the compound to a cell or organ. The treatment may be systemic or topical administration.

An effective amount is the amount of active ingredient that is administered in a single dose or multiple administrations necessary to achieve the desired pharmacological effect. The skilled physician can determine the effective dosage to the individual patient or can be determined by routine experimentation and titration known to the skilled clinician to treat the individual condition. However, unexpected clinical responses from a patient population to a pharmaceutical agent or composition may dictate unexpected changes or adjustments depending on the aspect of treatment, such as dosage, interval between drug administration and / or the manner of administration of the drug. The actual dosage and schedule are administered depending on whether the composition can be administered in combination with other drugs or on the basis of interspecific differences in pharmacokinetics, drug degradation and metabolism. Similarly, the amount may change for in vitro applications. When disclosed herein, the dosage range does not necessarily preclude the use of higher or lower doses of the ingredients, as is guaranteed in the specific application, unless otherwise indicated to the contrary.

The description of the pharmaceutical compositions provided herein includes pharmaceutical compositions suitable for administration to humans. One skilled in the art will appreciate based on the present disclosure that the composition is generally suitable for administration to any mammal or other animal. It will be appreciated that the manufacture of compositions suitable for administration to a variety of animals will be appreciated and ordinarily skilled artisans will be able to design and practice such modifications in routine experimentation based on pharmaceutical compositions for administration to humans.

Structure and properties of compounds

Naphtho furan compounds of formula I such as 2- (1-hydroxyethyl) -naphtho [2,3-b] furan-4,9-dione, 2- acetyl-7-chloro-naphtho [2,3 -b] furan-4,9-dione, 2-acetyl-7-fluoro-naphtho [2,3- b] furan-4,9-dione, 2- acetylnaphtho [2,3- b] furan Ethyl-naphtho [2,3-b] furan-4,9-dione can be obtained by reacting DMSO (dimethylsulfoxide), N-methylpyrrolidine, DMA (dimethylacetamide) (PEG 400 (polyethylene glycol 400), propylene glycol, Cremophor EL (polyethoxylated castor oil), Labrasol (caprylocaproyl macroroglyceride (polyoxyl glyceride)), M < / RTI > (vegetable oil PEG-6 (polyethylene glycol) ester) and capryl (propylene glycol caprylate). The naphthofuran compounds may be soluble in a wide range of polar organic solvents, such as certain halocarbons, such as chlorocarbons such as methylene chloride, esters, carboxylic acids such as ethyl acetate, acetic acid, ketones such as acetone, and alcohols such as methanol have. The naphthofuran compound was found to be soluble in methylene chloride and ethyl acetate.

Experimental studies described herein in which selective anti-tumor activity was found were achieved by administering the active compound of the pharmaceutical composition in small particle form to achieve specific pharmacokinetic exposure to selective anti-cancer activity focused on naphthofuran compounds . Conducting the observations discussed using the compounds similarly, the guitar naphthofuran, e.g., naphthofuran, can be used to achieve certain pharmacokinetic exposure in order to achieve selective anticancer activity upon administration in the form of small diameter particles It may be beneficial to change the pharmacokinetic profile. The pharmacokinetic profile of other naphthofuran administered as one or more different particle size distributions can be obtained by experiment.

Some other compounds that may exhibit an improvement in their pharmacokinetic profile and efficiency with a reduction in the specific size of the form administered to an animal, mammal, or human, as observed for the compound tested in the Examples, I, and salts and solvates thereof.

(I)

In formula (I), the indication (R ₁ ) _n indicates that the (R ₁ ) substituent is independently substituted at each available position along the benzene ring. For example, when n is 4, all four R ₁ substituents may be the same or may be different from one another. For example, each R ₁ is independently selected from hydrogen, halogen, fluorine, cyano, nitro, CF ₃ , OCF ₃ , alkyl, methyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, Wherein R is selected from the group consisting of hydrogen, alkyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heterocycle, substituted heterocycle, aryl, substituted aryl, OR _a , SR _a and NH ₂ . Alkyl may include, for example, a moiety having from 1 to 8 carbon atoms bonded by a single bond, and the alkenyl may be, for example, a straight or branched chain having from 2 to 8 carbon atoms And the alkynyl may have a moiety having from 2 to 8 carbon atoms bonded by, for example, one or more triple bonds. Substituents include moieties such as hydrogen, halogen, cyano, nitro, aryl, OR _a , SR _a and NH ₂ . For example, each (R ₁₎ is from the group consisting of hydrogen, methyl, F (fluorine), Cl (chlorine), Br (bromine), I (iodine), OH (hydroxyl) and NH ₂ (amine) Can be selected independently. For example, R ₃ can be hydrogen, halogen, fluorine, cyano, CF ₃ , OCF ₃ , alkyl, methyl, substituted alkyl, halogen-substituted alkyl, hydroxyl- Substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl, heterocycle, substituted heterocycle, aryl, substituted aryl, OR _a , SR _a and NR _b R _c . For example, R ₃ can be selected from the group consisting of methyl and C (R ₈ ) ₃ . Each (R ₈ ) may be independently selected from the group consisting of hydrogen, methyl, F (fluorine), Cl, Br, I, OH and NH ₂ . For example, two or less of the independently selected (R ₁ ) substituents and (R ₈ ) substituents may be selected to be F (fluorine), with the remainder selected to be hydrogen.

In some embodiments, the compound of formula I is selected from the group consisting of 2- (1-hydroxyethyl) -naphtho [2,3-b] furan-4,9-dione, 2- acetyl- 2,3-b] furan-4,9-dione, 2-acetylnaphtho [2,3-b] Furan-4,9-dione, 2-ethyl-naphtho [2,3-b] furan-4,9-dione, its enantiomers, diastereoisomers, tautomers and salts or solvates . For example, each (R ₁₎ may be selected from hydrogen, R ₃ is the compound of formula (I) so that the 2-acetyl-naphtho [2,3-b] furan-4,9-dione is selected as the methyl . For example, each R _a is independently selected from hydrogen, alkyl, substituted alkyl, alkenyl, substituted alkenyl, alkynyl, substituted alkynyl, cycloalkyl, substituted cycloalkyl, cycloalkenyl, substituted cycloalkenyl , Heterocycle, substituted heterocycle, aryl, and substituted aryl. For example, each R _b and R _c may be independently selected from the group consisting of hydrogen, alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, heterocycle, substituted heterocycle, aryl, and substituted aryl have. Alternatively, R < _b & gt _; and R < _c > together with the N to which they are attached may form a heterocycle or a substituted heterocycle.

Polymorphism

The naphthofuran compounds of the present invention include polymorphs. In some embodiments, the polymorphism is a polymorph of the compound according to formula (I). In some embodiments, the polymorphism is a polymorph of Compound 1. For example, in some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2 < RTI ID = 0.0 > , 3-b] furan-4,9-dione. Such polymorphs are referred to herein as "crystal form 1 "," form 1 "or" XRPD1 ", and these terms are used interchangeably. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] pyrimidine which is characterized by an X-ray diffraction pattern substantially similar to that specified in Figure 2 of WO 2011/116398 and WO 2011/116399 ] Furan-4,9-dione. Such polymorphs are referred to herein as "crystal form 2 "," form 2 "or" XRPD2 ", and such terms are used interchangeably. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b < RTI ID = 0.0 > ] Furan-4,9-dione. Such polymorphs are referred to herein as "crystal form 3", "form 3" or "XRPD3", and these terms are used interchangeably.

For example, in some embodiments, the polymorphism is an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.4, 11.9, 14.1, 14.5, 17.3, 21.0, 22.2, 24.0, 26.0, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H, 9H, 9H, 9H, or 9H, characterized by an X-ray diffraction pattern comprising at least one peak at at least about 10.2, 11.9, 14.1, 14.5, 17.3, -Naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 10.2 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 11.9 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.1 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 14.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 17.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 22.2 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.1 [ It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 10.2 degrees two theta, a peak at least about 11.9 degrees two theta, a peak at least about 14.1 degrees two theta, a peak at least about 14.5 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at at least about 22.2 DEG 2 & -Naphtho [2,3-b] furan-4,9-dione.

For example, in some embodiments, the polymorphism is an X that comprises at least one peak at at least about 7.5, 9.9, 11.4, 12.3, 15.0, 23.0, 23.3, 24.1, 24.6, 25.0, 26.1, Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, which is characterized by a line diffraction pattern. In some embodiments, the polymorphs are 2-acetyl-4H, which is characterized by an X-ray diffraction pattern comprising at least one peak at at least about 7.5, 9.9, 12.3, 15, 23.0, 23.3, 24.6 and / , 9H-naphtho [2,3-b] furan-4,9-dione. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 7.5 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 9.9 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 12.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 15 & It is a polymorph of Dion. In some embodiments, the polymorphism is a 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione which is characterized by an X-ray diffraction pattern comprising peaks at at least about 23 [ It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 23.3 & It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 24.6 degrees two- It is a polymorph of Dion. In some embodiments, the polymorphs are 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern comprising peaks at at least about 28.4 degrees two- It is a polymorph of Dion. In some embodiments, the polymorph has a peak at at least about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 12.3 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at least about 23.3 DEG 2 &thetas; a peak at least about 24.6 DEG 2 DEG and at least about 28.4 DEG 2 theta and any combination thereof Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione.

Crystal Form 1 is detected in various solvents and conditions, but has low antitumor activity (WO 2011/116398 and WO 2011/116399, FIG. 8). In the study shown in FIG. 9 of WO 2011/116398 and WO 2011/116399, immunosuppressed mice with established subcutaneous FaDu human head and neck cancer were treated with the indicated doses of passive milled Compound 1 or vehicle control with crystalline Form 1 in oral po). Compound 1 was formulated in Gel Lucir ™. All regimens were administered qd daily. Tumor size was assessed periodically during treatment.

Crystalline form 2 was surprisingly obtained in the presence of impurities, and these polymorphs were found to exhibit effective antitumor activity (WO 2011/116398 and WO 2011/116399, FIG. 9). In the study shown in FIG. 9 of WO 2011/116398 and WO 2011/116399, immunosuppressed mice with established subcutaneous FaDu human head and neck cancer were given the synthetic method described in Figures 5A and 5B of WO 2011/116398 and WO 2011/116399 100 mg / kg of undifferentiated Compound 1 (first crop) or a vehicle control produced in Example 1 is orally administered (po). Compound 1 was formulated in Gel Lucir ™. All regimens were administered qd daily. Tumor size was assessed periodically during treatment. Form 2 was successfully manufactured by the current Good Manufacturing Practice (cGMP) process and approved for use in clinical trials by the FDA and the Canadian Department of Health. Form 2 shows the favorable pharmacokinetics of antitumor activity in cancer patients (WO 2011/116398 and WO 2011/116399, FIG. 11), indicating stability and a strong indication.

Crystal Form 3 was similar to Form 1, but shared different X-ray powder diffraction (XRPD) patterns and showed much different crystal habits than Form 1 (see Figures 7A and 7B of WO 2011/116398 and WO 2011/116399 7B). Form 3 may be produced only from Form 1 using the specially designed slurry procelse described herein. Form 3 has been shown to exhibit effective antitumor activity (WO 2011/116398 and WO 2011/116399, FIG. 10). In the study presented in FIG. 10 of WO 2011/116398 and WO 2011/116399, immunosuppressed mice with established subcutaneous FaDu human head and neck cancer were treated with 200 mg / kg of Compound 1 with passively pulverized crystal form 1 or 3 The vehicle control was administered orally (po). Compound 1 was formulated in gel lucer. All regimens were administered qd daily. Tumor size was assessed periodically during treatment. These polymorphisms were successfully produced by the cGMP process and approved for use in clinical trials by the FDA and the Canadian Department of Health. Form 3 shows the desired pharmacokinetics of antitumor activity in cancer patients (WO 2011/116398 and WO 2011/116399, Figure 12), safety and strong indication.

Synthetic methods for generating crystal form 2 are shown in Figures 5A-5B of WO 2011/116398 and WO 2011/116399. Briefly, charged 3-buten-2-one (451.2 g) is added to a 2 L three neck round bottom flask equipped with a mechanical stirrer, thermometer and addition funnel. Bromine (936.0 g) was added to the addition funnel. After cooling the contents in the flask to -5 ° C, bromine is added dropwise to the flask over 30 minutes while maintaining the temperature at -5 ° C with vigorous stirring. The mixture is stirred at -5 [deg.] C for an additional 15 minutes and then divided into four equal parts. Each portion of the mixture with tetrahydrofuran (2,133.6 g) is added to a 22 L four-neck round bottom flask equipped with a mechanical stirrer, thermometer and addition funnel. DBU (1,3-diazabicyclo [5.4.0] undec-7-ene, 222.9 g) is added to the addition funnel. The DBU is added dropwise to the flask over 30 minutes while maintaining the temperature at 0 占 폚 -5 占 폚 under vigorous stirring. The mixture was stirred at < RTI ID = 0.0 > 0 C < / RTI > Then, 2-hydroxy-1,4-naphthoquinone (231 g) is added to the flask. Additional DBU (246.0 g) is added to the addition funnel and added dropwise to the mixture in the flask at such a rate that the temperature of the reaction mixture does not exceed 40 ° C. After the addition of DBU is complete, the resulting mixture is stirred overnight at room temperature and a sample of the reaction mixture is run for HPLC analysis. Water (10.8 L) is added to the reaction mixture, the resulting mixture is cooled to 0 to -3 DEG C for at least 30 minutes, and then filtered by a vacuum filter. The filtered solid is rinsed successively with 5% aqueous sodium bicarbonate (3 L), water (3 L), 1% aqueous acetic acid (3 L) and twice with ethanol (2 L 1 L). Store the rinse solids and fuse together from the other batches. The combined crude product (28.73 kg) is added to a 500 gallon vessel equipped with a mechanical stirrer, thermometer and condenser along with ethyl acetate (811.7 kg). Under a nitrogen atmosphere, the mixture is heated to reflux (72 ° C) for 2 hours and then filtered through a 10 micron cartridge filter containing an activated carbon layer to remove insolubles. Rinse the vessel, transfer line and filter with fresh hot ethyl acetate (10 kg). The combined filtrates are cooled to 0-5 < 0 > C, held at this temperature for 2 hours, and filtered through a 20-inch Buchner filter. The filtered solid product is rinsed with 0-5 ° C ethyl acetate (5.7 kg) and dried under vacuum at 40 ° C to constant weight. The remaining filtrate was reduced to a volume of up to 63% by evaporation and the crystallization process was repeated again to produce a second crop of product which was also dried as the first crop of product under the same conditions. Both crops obtained are in crystalline form 2. The first crop (0.5 kg) produced had 99.5% purity by HPLC (~ 95% by NMR). The resulting second crop (1.09 kg) had 98.9% purity by HPLC (~ 90% by NMR).

A synthesis process for producing crystal form 3 is shown in Figures 6A-6D of WO 2011/116398 and WO 2011/116399. The steps are briefly described herein. Step 1: 3-buten-2-one (methyl vinyl ketone, MVK) is brominated using bromine. No additional solvent is used. The intermediate 3,4-dibromobutan-2-one was dissolved in tetrahydrofuran (THF) and reacted with 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU) Bromo-3-buten-2-one, which is an intermediate of the 3-bromo-3-butene. When the reaction is complete, 2-hydroxy-1,4-naphthoquinone (HNQ) is added. The second portion of the DBU is added and the mixture is exposed to air. The reaction is quenched with water and the solid is collected by filtration. The solids are washed with aqueous sodium bicarbonate, aqueous acetic acid, water and ethanol. The product is slurried in ethanol and the solid is collected and separated. Step 2: The residual amount of 2-acetyl-2, 3-dihydronaphtho [2, 3-b] furan- 2,3-b] furan-4,9-dione is oxidized to compound 1 using activated manganese dioxide in toluene. The mixture is filtered through a cake of charcoal and celite. The filtrate is concentrated to precipitate the product, which is filtered and dried. Step 3: The solid is slurried in ethyl acetate (25 ml / g purified Compound 1) at 75-80 < 0 > C for about 5 hours, collected by filtration and dried. Compound 1 produced in this manner is in crystalline form 3. Compound 1 produced in this manner without slurry process produced crystal form 1.

Effect of compound particle size distribution on plasma drug concentration and selective anti-tumor activity

In the prior art for the present invention, the microparticles of Compound 1 were not produced and / or evaluated. Conventional studies have shown that Compound 1 is equally toxic to normal and cancer cells, and antitumor activity has not been observed in animal models. The studies presented here demonstrate that particle size reduction of Compound 1 not only improves bioavailability, but also increases selective antitumor activity without the sign of toxicity. Improvement in bioavailability is unexpected as it increases the exposure of compound 1 to cancer cells and normal cells as well. A mechanism for selective enhancement of anticancer activity without improving toxicity to normal cells is not known. In this study, this improvement in bioavailability of Compound 1 was found to be maximal when D ₅₀ (i.e., the median point of the particle size distribution dividing the distribution into two identical portions) was about 20 μm. However, additional experiments were conducted when the D ₅₀ value was about 2 탆. Microparticles of Compound 1 with a two-micron D ₅ may even represent an improvement in the pharmacokinetic exposure as compared to particles having a D ₅₀ of 20 microns jinyeotda surprisingly enhanced anti-tumor activity. In a further experiment, but the nanoparticles of the compound 1 having a _{D 50 (D 50 = 110.4 ㎚} ) of about 100 ㎚ produced, surprisingly reduce the anti-tumor activity was observed with these particle size of one of the compounds. Thus, in a preferred embodiment, a composition containing particles of Compound 1, for example microparticles, has a D ₅₀ of less than or equal to 20 microns of 0.2 microns and has surprisingly effective antineoplastic activity without an increase in cytotoxicity to normal cells.

The antitumor activity of the particles of Compound 1 having different particle size ranges is illustrated in FIG. 15 of WO 2011/116398 and WO 2011/116399, and pharmacokinetic data for particles of Compound 1 having different particle size ranges can be found in WO 2011 / 116398 and WO < RTI ID = 0.0 > 2011/63999 < / RTI > In the experiments shown in WO 2011/116398 and WO 2011/116399, an immunosuppressed mouse with established subcutaneous FaDu human head and neck cancer is administered orally (po) in a given amount of Compound 1 or vehicle control with the indicated particle size, . All regimens were administered qd daily. Tumor size was assessed periodically.

It has been found that administering a naphthofuran compound in the form of particles with a defined particle size, for example, a reduced particle size, improves in vivo plasma drug concentration. In the present application, unless the context clearly indicates otherwise, the terms "size" and "diameter" will be used to describe the particles alternately. It should be understood that the use of the term " diameter " does not necessarily imply that the particles have a perfect or approximate spherical shape. For example, "diameter" can be used as an approximation of the particle size, for example, the diameter of a sphere of equivalent volume to a non-spherical particle.

In surprising results, it has been found that administration as a naphthofuran compound particle, e.g., small particle, of a defined particle size distribution in a pharmaceutical composition results in selective antitumor activity. For example, compounds administered as particles with a median particle size of 20 microns (i.e., microns, alternately used herein) exhibit relatively weak, but potent (selective antitumor activity) in the mouse xenograft model. By comparison, particles of 150 microns (microns) did not show efficacy. The discovery that administration of naphthofuran compounds in the form of smaller particles may result in selective antitumor activity is surprising and can not be explained on the basis of solubility or bioavailability alone improvement. That is, generally, improved solubility is associated with antitumor activity, as well as increased drug oral bioavailability that can improve toxicity to normal cells. As discussed above, a naphthofuran compound may be equivalent toxicity to tumor cells and normal cells if exposure is not carried out under established conditions as described in WO 2009/036099 and WO 2009/036101.

In an additional surprising result, administration of further reduced-sized particles of naphthofuran compound particles in the pharmaceutical composition resulted in greatly improved anti-tumor activity, but resulted in an almost unchanged pharmacokinetic profile, i. . For example, compounds administered as particles with a median particle size of 2 [mu] m (microns) showed significantly improved efficacy in the mouse xenograft model. Compared to particles of 20 μm, particles of 2 μm exhibit greatly improved efficacy but exhibit a very similar pharmacokinetic profile. In other words, such improved efficacy is independent of the pharmacokinetic profile, bioavailability. Because of the poor solubility of compounds, improved efficacy is generally associated with increased drug oral bioavailability, which is quite surprising.

Therefore, the observed improvement in selective antitumor activity is surprising and unexpected. The present invention provides naphthofuran compounds, for example, particles or particles of a compound of Formula I, having activity, i. E. Having potency or selective anti-tumor activity. The active particles or particles have a defined particle size, for example, about 200, about 150, about 100, about 40, or about 20, about 10, about 5, about 4, About 2 microns, about 1 micron, about 0.5 microns, about 0.2 microns, or about 0.1 microns or less. Particles or particles larger than the defined particle size are less active or inert than the particles described herein.

Thus, administration of another compound or naphthofuran compound according to formula I in the form of smaller particles may result in an improvement in its selective antitumor activity. The use of particles of a compound according to formula I having a defined particle size distribution in administration can permit the establishment of the desired selective anti-tumor activity. For example, the use of naphthofuran compound particles, e.g., smaller particles, with a defined particle size distribution can result in higher blood concentration and selective anti-tumor activity for a relatively short but short period of time. An additional reduced particle size of the compound can lead to an unaltered plasma concentration of the compound with greatly improved efficacy.

In the present application, unless otherwise indicated, the terms "plasma concentration", "blood concentration" and "blood concentration" are used interchangeably. The term "neoplasm" can be used to describe a cell that exhibits an abnormal pattern of growth. Such neoplasms include benign and malignant tumors such as solid tumors, as well as other cell growth disorders such as leukemia that do not have a defined shape and are not limited to a particular part of the human or animal body. Thus, "neoplasm" includes both cancerous and non-cancerous neoplastic cells and tissues. The terms "tumor" and "cancer ", unless the context clearly dictates otherwise, or refer to a particular study or experiment, are intended to encompass all types of neoplasia, including but not limited to certain parts of the human or animal body As used herein. It should be understood, however, that the more limited term "solid tumor" does not have a definite form and does not include cell growth disorders such as leukemia, which are not limited to a particular region of the human or animal body.

The neoplasm may or may not exhibit one or more of the following characteristics: solid form (solid tumors), malignancy, metastasis or Stat3 pathway activity. The neoplasm may include, for example, cancer stem cells. The neoplasm may include, for example, carcinoma, sarcoma, adenocarcinoma, lymphoma or hematologic malignancies.

Absorption is defined as the process of transferring a drug from the site of administration to the site of measurement in the body. [M. Rowland, TN Tozer (1995) Clinical pharmacokinetics: Concepts and applications. Lippincott Williams & Wilkins]. Oral drug absorption is often referred to as drug delivery across the mucous membrane of the small intestine, since the membrane is considered to be a rate limiting step for permeation of the membrane. U. Fagerholm & H. Lennernaes (1995) Experimental estimation of the effective unstirred water layer thickness in the human jejunum, and its importance in oral drug absorption, Eur J Pharm Sci 3: 247-253; MB Lande, JM Donovan & ML Zeidel (1995), The relationship between membrane fluidity and permeabilities to water, solutes, ammonia and protons, J Gen Physiol 106: 67-84. Permeability is a general term that indicates how easily a drug passes through a membrane. The specific permeability characteristics of the drug depend on its physico-chemical properties including lipophilicity, charge, size and polarity surface area. Rowland & Tozer 1995; CA Lipinski, F. Lombardo, and BW Dominy & PJ Feeney (2001), Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings, Adv Drug Deliv Rev 46: 3-26. The rate of absorption depends on the permeability of the drug, the surface area of the membrane and the concentration gradient to the membrane. Concentration gradient is the driving force for passive diffusion, the most common mechanism for drug membrane transport. In the case of oral administration, the drug is mainly absorbed by the small intestine. The human small intestine is about 5-8 meters long and has a total surface area of almost 200 square meters while the small intestine of the mouse is only about 10-20 cm long. Thus, while the permeability of a drug having a larger particle size is smaller than that of a drug having a smaller particle size, a drug having a larger particle size is more effective than a drug having a smaller particle size in a mouse It can be predicted that it can have the absorption rate of < RTI ID = 0.0 >

For example, it is preferable to use a material having a thickness of about 200 占 퐉, 150 占 퐉, 100 占 퐉, 80 占 퐉, 60 占 퐉, 40 占 퐉, 20 占 퐉, 10 占 퐉, 5 占 퐉, 4 占 퐉, 3 占 퐉, 2 占 퐉, The distribution of the particle size of the compounds according to formula I with the median diameter of the compounds of the invention may be expected to result in selective antitumor activity upon administration, for example, to pharmaceutical preparations for the treatment of cancer or tumors. For example, the distribution of particle sizes can cause the particles to have a median diameter of about 0.02 μm to about 5 μm, or about 0.2 μm to about 4 μm. For example, the distribution of particle sizes may allow the particles to have a median diameter of about 5 占 퐉 or less, a ratio of the average diameter to a median diameter of about 2 or less, and a median diameter to median diameter of about 0.25 or more.

The term "particle" can refer to an aggregate of a compound of formula (I). The term "average" can refer to the sum of the sizes of all particles divided by the total number of particles. The term "center" may, for example, refer to a half of the particles having a larger diameter and a half of the particles having a smaller diameter. The term "mode" may represent the most commonly occurring particle size value. The term "cumulative sum" can refer to all particles.

로서 정의될 수 있으며, 여기서 V_p는 입자의 부피이며, A_p는 입자의 표면적이다. 구체는 Ψ = 1의 구형도를 가지며, 입자의 구형도가 1(unity)에 근접할수록 입자의 형상은 구체의 근사치에 더 가깝다. 비교에 의하면, 4면체는 약 0.671의 구형도를 가지며, 정육면체는 약 0.806의 구형도를 가지며, 8면체는 약 0.846의 구형도를 가지며, 12면체는 약 0.910의 구형도를 가지며, 20면체는 약 0.939의 구형도를 갖는다. 구체의 형태가 주어진 부피에 대한 표면적을 최소로 하므로, 거의 구체인 입자는 덜 구체에 가까운 동일한 부피의 입자보다 더 느리게 흡수될 것으로 예상될 수 있다. 구체의 세트의 평균 구형도는

로서 정의될 수 있으며, 여기서 ΣV_p는 모든 입자의 총 부피이며, ΣA_p는 모든 입자의 총 표면적이다. 예를 들면, 투여되는 화학식 I에 의한 화합물의 입자는 약 0.8 이상의 평균 구형도 또는 약 0.9 이상의 평균 구형도를 가질 수 있다.The selective antitumor activity achieved by administration of the naphthofuran compound particles may depend on the size distribution of the particles, for example the diameter or the volume of the particles representing these volumes, as well as the distribution of the shape and shape of the particles. For example, a set of particles having a needle-like shape may result in a different pharmacokinetic profile than a set of particles having a spherical shape. Thus, a method of measuring the shape and shape distribution of a particle to be administered and / or generating a particle having a predetermined shape and shape distribution, for example, an approximately uniform shape, for example, an approximation of a sphere May be preferred. For example, the spherical degree Ψ of the particle is

, Where V _p is the volume of the particle and A _p is the surface area of the particle. The sphere has a sphericity of Ψ = 1, and the closer the sphericity of the particle is to 1 (unity), the closer the particle's shape is to the sphere's approximation. According to the comparison, the tetrahedron has a sphericity of about 0.671, the cube has a sphericity of about 0.806, the octahedron has a sphericity of about 0.846, the dodecahedron has a sphericity of about 0.910, And has a sphericity of about 0.939. Since the shape of the sphere minimizes the surface area for a given volume, it can be expected that nearly spherical particles will be absorbed more slowly than particles of the same volume that are closer to less spherical. The average sphericity of the set of spheres is

Where ΣV _p is the total volume of all particles, and ΣA _p is the total surface area of all particles. For example, the particles of the compound according to formula I administered may have an average sphericity of at least about 0.8 or an average sphericity of at least about 0.9.

The size, size distribution, shape, shape distribution and factors, such as surface roughness or particle irregularity, can affect the average specific surface area of a set of Compound 1 particles administered in a pharmaceutical formulation. The average specific surface area can be defined as ΣA _p / Σm _p , where ΣA _p is the total surface area of the particles and Σm _p is the total mass of the particles. The larger the average specific surface area of the particles, the faster the expected dissolution of the particles.

The particles of the compounds according to formula I in the pharmaceutical preparations may contain naphthofuran compounds in crystalline states across different particles or within the same particle. The crystalline state can comprise one or more polymorphs across different particles or within the same particle. As will be appreciated by those skilled in the art, the rate of dissolution of the particles is expected to be influenced by the state of the material in the compound particles, for example, regardless of whether they are the first polymorph or the second polymorph.

One or more of the various techniques can be applied to determine the size and / or size distribution of the particles of a compound according to formula I in a pharmaceutical formulation. For example, sieve analysis, optical microscopy, electron microscopy, electrical resistivity, settling time, laser diffraction and / or acoustic spectroscopy can be applied. Some or all of these techniques, or variations thereof, may be applied to determine the shape, shape distribution and / or specific surface area of the naphthofuran compound particles in the pharmaceutical preparation. The BET isotherm and / or air permeability specificity surface technique can be applied to determine the specific surface area of a particle of a compound according to formula I in a pharmaceutical formulation.

Naphthofuran  Process for the production of compounds

WO 2009/036099 and WO 2009/036101 disclose a process for preparing a naphthofuran compound of the formula II as follows:

In this process, 3-bromo-3-buten-2-one (4-3) was reacted with 2-hydroxy-1,4-naphthoquinone (4-4) -Dihydronaphtho [2,3-b] furan-4,9-dione (4-5). 2,3-b] furan-4,9-dione (4-5) is oxidized by oxygen from open air to form naphtho [2,3-b] furan- 9-dione (4-6). This process produces naphtho [2,3-b] furan-4,9-dione. However, during further production of the compounds, this process has been determined to produce a significant variety of impurities that interfere with potential clinical applications of these compounds. In some embodiments, one of the impurities is 2,3-dihydronaphtho [2,3-b] furan-4,9-dione (4-5).

In one embodiment, the present invention provides an improved method of producing naphthofuran. The improved method produces substantially pure naphtho furan with minimal impurities. As used herein, the term "substantially pure" refers to a preparation comprising at least about 80% of a compound of the present invention as measured by% area HPLC. In some embodiments, the naphthofuran is naphtho [2,3-b] furan-4,9-dione and its related compound (4-6).

In some embodiments, the method of the present invention comprises one of the methods set forth in the working examples provided herein. In some embodiments, the method comprises one or more of the methods set forth in Examples 1, 2 and / or 5 provided herein.

In some embodiments, the process of the present invention comprises reacting 3-bromo-3-buten-2-one (4-3) and 2-hydroxy-1,4-naphthoquinone (4-4) Lt; / RTI > with an oxidizing agent. In a further embodiment, the oxidizing agent is manganese dioxide (MnO _2). In a further embodiment, the crude product is separated prior to oxidation. In some embodiments, the first solvent is toluene or chloroform.

In some embodiments, the method of the present invention further comprises treating the aged oxidation mixture with a charcoal to remove certain impurities. In a further embodiment, the aged oxidation mixture is filtered through a pad of activated carbon. In a further embodiment, the mixture is filtered at about < RTI ID = 0.0 > 100 C. < / RTI >

In some embodiments, the method of the present invention further comprises crystallizing the product from the filtrate. In a further embodiment, the filtrate is concentrated by evaporation and cooled to crystallize the product.

In some embodiments, the process of the present invention further comprises recrystallizing the product to a second solvent. In a further embodiment, the second solvent is ethyl acetate.

In an alternative embodiment, the method of the present invention further comprises slurring the crystallized product from the first solvent in a second solvent, heating the slurry, and cooling the slurry. In a further embodiment, the second solvent is ethyl acetate. In some embodiments, the product is slurried and heated to only partially dissolve. In a further embodiment, the volume of the second solvent used to slurify the product is about 1/10, 1/5, 1/4, 1/3, 1 or 2 times the volume for complete dissolution of the product in the heated state / 2 or 2/3.

The present invention also provides naphthofuran compounds produced by the process of the present invention. In some embodiments, the naphthofuran compound is selected from the group consisting of 2- (1-hydroxyethyl) -naphtho [2,3-b] furan-4,9-dione, 2- acetyl- 2,3-b] furan-4,9-dione, 2-acetylnaphtho [2,3-b] Furan-4,9-dione, 2-ethyl-naphtho [2,3-b] furan-4,9-dione, mono - [1- (4,9-dioxo-3a, 2-yl) -vinyl] ester, phosphoric acid 1- (4,9-dioxo-3a, 4,9,9a-tetrahydro-naphtho [2 , 3-b] furan-2-yl) -vinyl ester dimethyl ester, its enantiomers, diastereoisomers, tautomers and salts or solvates thereof. In a further embodiment, the naphthofuran compound is prepared by coupling 2-hydroxy-1,4-naphthoquinone (4-4) and 3-bromo-3-buten- And reacting the separated crude product with manganese dioxide in the presence of toluene. In a further embodiment, the method further comprises filtering the aged reaction mixture through an activated carbon pad.

In another embodiment, the present invention provides a substantially pure naphthofuran compound.

In some embodiments, the present invention provides a process for the preparation of 2- (1-hydroxyethyl) -naphtho [2,3-b] furan-4,9-dione, 2- acetyl- 2,3-b] furan-4,9-dione, 2-acetyl-7-fluoro-naphtho [ 4,9-dione, 2-ethyl-naphtho [2,3-b] furan-4,9-dione, mono - [1- (4,9-dioxo-3a, 2-yl) -vinyl] ester, phosphoric acid 1- (4,9-dioxo-3a, 4,9,9a-tetrahydro-naphtho [2,3 -b] furan-2-yl) -vinyl ester dimethyl ester, its enantiomers, diastereoisomers, tautomers and salts or solvates thereof.

In some embodiments, the invention provides substantially pure compounds of formula II:

(II)

(II)

In the above formula, each R < ₁ > is independently H, Cl or F; n is 0, 1, 2, 3 or 4;

As used herein, "substantially pure" refers to a purity of at least about 80%. In some embodiments, the purity of the compounds of the invention has a purity of at least about 85%, about 90%, about 95%, or about 99%. In a further embodiment, the purity of the compounds of the invention has a purity of at least about 99.5% or about 99.8%. In a further embodiment, the purity of the compounds of the present invention has a purity of at least about 99.85%, about 99.90%, about 99.94%, about 99.95%, or about 99.99%. In some embodiments, a compound of the present invention is 2- (1-hydroxyethyl) -naphtho [2,3-b] furan-4,9-dione, 2- acetyl- 7- chloronaphtho [2,3 -b] furan-4,9-dione, 2-acetyl-7-fluoro-naphtho [2,3- b] furan-4,9-dione, 2- acetylnaphtho [2,3- b] furan 4,9-dione, 2-ethyl-naphtho [2,3-b] furan-4,9-dione, mono - [1- (4,9-dioxo-3a, 2-yl) -vinyl] ester, phosphoric acid 1- (4,9-dioxo-3a, 4,9,9a-tetrahydro-naphtho [2, 3-b] furan-2-yl) -vinyl ester dimethyl ester, its enantiomers, diastereoisomers, tautomers and salts or solvates thereof. In some embodiments, the compounds of the invention are multimeric. In some embodiments, the compounds of the present invention are polymorphs of the compounds according to formula (I). In some embodiments, the compounds of the invention are polymorphs of Compound 1.

Conventional impurities that may be present in the compounds of the present invention include at least one selected from the group consisting of byproducts, isomers, intermediates and solvents. In some embodiments, the impurities that may be present in the compounds of the invention are about 10%, about 8%, about 5%, about 2%, or about 1% or less of the compound of Formula II. In a further embodiment, the impurities that may be present in the compounds of the invention are about 0.5%, about 0.2%, about 0.15%, or about 0.1% or less, relative to the compound of Formula II. In a further embodiment, the impurities present in the compounds of the invention are about 0.05%, about 0.02%, or about 0.01% or less, relative to the compound of formula (II). In some embodiments, a substantially pure compound of formula II is present in an amount of about 500, 200, 100, 50, 20, 10, 5, 2, 1, 0.5, 0.2, 0.15, By-products or by-products.

In some embodiments, the impurity is selected from the group consisting of 2-acetyl-2,3-dihydronaphtho [2,3-b] furan-4,9-dione, 2,6-diacetyl- Furan-4,9-dione, 2,7-diacetyl-naphtho [2,3-b] furan-4,9-dione, 3-acetyl-naphtho [2,3- b] 2,3-b] furan-4,9-dione, naphtho [2,3-b] furan-4,9-dione, naphtho [2,3- b] furan- Diol and 1- (4,9-dihydroxy-naphtho [2,3-b] furan-2-yl) -ethanone.

In some embodiments, the impurity comprises manganese (Mn).

The purity of the compounds of the present invention can be measured using various devices. In some embodiments, purity is determined using HPLC (high performance liquid chromatography). In some embodiments, purity is measured using NMR (nuclear magnetic resonance). In a further embodiment, purity is determined using HPLC and NMR.

These very pure compositions containing Compound 1 exhibit a greatly improved stability profile in animal experiments compared to less pure compositions containing Compound 1. [ No signs of any harmful effects of highly pure Compound 1 were observed in mice. In addition, these very pure compositions containing Compound 1 were tested in patients, demonstrating excellent stability. For example, FIG. 13 of WO 2011/116398 and WO 2011/116399 illustrate the observed toxicity using a composition having about 90% purity for compound 1, see WO 011/116398 and WO 2011/116399, FIG. 14 Illustrate that very pure compositions having a purity of about 95% or greater with respect to Compound 1 are safe and effective. In the study shown in FIG. 13 of WO 2011/116398 and WO 2011/116399, an amount of compound given to an immunosuppressed mouse with established subcutaneous FaDu human head and neck cancer (upper panel) or MDA-231 human breast cancer (lower panel) 1 < / RTI > or vehicle control was administered orally (po). Compound 1 was formulated in Gel Lucir ™. All regimens were administered qd daily. Body weights were evaluated periodically during treatment. Each view represents the mean ± SEM of 8 tumors. Significant toxicity was observed in about 90% pure Compound 1. Since a total of four mice died during treatment in the first experiment (upper panel) (two mice on day 16, two mice on day 19, one mouse on day 23), their body weight was not included in the post-mortem plot. Since a total of 3 mice died during treatment in the second experiment (lower panel) (1 day on day 14, 2 days on day 21), their body weight was not included in their post-mortem plot. In the study shown in FIG. 14 of WO 2011/116398 and WO 2011/116399, immunosuppressed mice with established subcutaneous FaDu human head and neck cancer (upper panel) or MDA-231 human breast cancer (lower panel) 1 < / RTI > or vehicle control was administered orally (po). Compound 1 was formulated in Gel Lucir ™. All regimens were administered qd daily. Body weights were evaluated periodically during treatment. Each view represents the mean ± SEM of 8 tumors. Compound 1 with higher purity was well tolerated and showed no signs of toxicity. All mice remained healthy during the treatment in both experiments. In the Phase I experiment, the dose of Compound 1 was expanded from 20 mg to 2,000 mg / day, and the maximum tolerated dose (MTD) was not reached. No dose-limiting toxicity was observed. The patient tolerated Compound 1 very well without a drug-induced harmful effect, which is a striking contrast to cancer chemotherapeutic agents. The clinical safety profile of the substantially pure composition of Compound 1 is the highest of all oncological drugs of historical interest.

Pharmaceutical formulation

Certain excipients or enhancers have been found to improve the oral bioavailability of particles of a compound according to formula I of a given particle size distribution in a pharmaceutical formulation. For example, the addition of the pharmaceutical compatibility excipient Gelusir ™ 44/14 (polyethylene glycol glyceryl laurate made by Gatefosse) increases the bioavailability of Compound 1 with a median particle size of less than about 20 microns . Examples of other excipients that can be used to enhance or control oral bioavailability include surfactants such as TWEEN 80 ™ or Tween 20 ™ (polysorbate, ie, polyoxyethylene sorbitan monolaurate) or specific lipids, For example, phosphatidylcholine, such as dimyristoylphosphatidylcholine (DMPC). Surfactants include those that are both amphoteric and contain both hydrophobic and hydrophilic groups. Other excipients include, for example, glycerol esters of fatty acids, glycerol esters of saturated fatty acids, glycerol esters of saturated fatty acids having 8 to 18 carbons, glyceryl laurate, polyethylene glycol, polyoxyethylene sorbitan alkylate, Cellulose derivatives such as microcrystalline cellulose and carboxymethylcellulose (CMC), as well as lipids such as sterols, such as cholesterol. Other excipients include antioxidants such as vitamin E, for example. Other excipients and additional ingredients may be included in the pharmaceutical preparations according to the invention as will be appreciated by those skilled in the art. For example, it is possible to use other active agents, standard vehicles, carriers, liquid carriers, saline solutions, aqueous solutions, diluents, surface active agents, dispersants, inert diluents, granulating and disintegrating agents, lubricants, lubricants, A preservative, a physiologically degradable composition such as gelatin, an aqueous vehicle and a solvent, an oily vehicle and a solvent, a suspending agent, a dispersing or wetting agent, a suspending agent, an emulsifier, an emollient, a buffering agent, a salt, an agitator, a gelatin, Such as antioxidants, antibiotics, antifungal agents, stabilizers, water, glycols, oils, alcohols, crystallization retarders (for example retarding the crystallization of sugars), starches, sugars, sucrose, surfactants, Materials that increase the solubility of hydroxy alcohols, such as glycerol or sorbitol, pharmaceutically acceptable polymers or hydrophobic materials, and other ingredients. Additional materials or materials suitable for addition will depend upon the dosage form (e. G., Injectable solutions, capsules or pills) as will be appreciated by those skilled in the art.

The compounds according to formula I of the present invention may be formulated into "pharmaceutical compositions." Embodiments of the present invention include various forms of administration including, for example, compounds that may be useful in treating patients. For example, oral dosage forms may be in the form of tablets, pills, capsules (hard or soft), sugar-coated tablets, powders, granules, suspensions (for example in an aqueous or oily vehicle), liquid (for example in an aqueous or oily vehicle) , Cachets, troches, lozenges, syrups, elixirs, emulsions, potions, oil-in-water emulsions or water-in-oil emulsions. Due to the ease of administration, tablets and capsules may exhibit desirable oral administration. Solid oral dosage forms can be administered orally, by standard techniques. For example, nasal and other mucosal spray formulations (e.g., inhalable formulations) may contain the active compound and a purified aqueous solution of an antiseptic and an isotonic agent. The formulation is preferably adjusted to a pH and isotonic state that is compatible with the nasal cavity or other mucosa. Alternatively, they may be present in the form of finely divided solid powders suspended in a gas carrier of an inhaler or aerosol. The formulation may be delivered by any suitable means or method, for example, by a nebulizer, atomizer, metered dose inhaler, and the like. For example, the pharmaceutical compositions according to the invention may be administered topically, for example in the form of ointments, creams or suppositories. For example, the pharmaceutical composition according to the present invention can be administered by injecting an injection. Thus, dosage forms according to the present invention may have, for example, solid, semi-solid, liquid or gaseous forms. Suitable dosage forms include, but are not limited to, oral, rectal, sublingual, mucosal, nasal, ocular, subcutaneous, intramuscular, intravenous, parenteral, transdermal, spinal, intrathecal, intraarticular, subarachnoid, And other forms of administration for systemic delivery of the active ingredient. The active ingredient, for example, a compound according to Formula I, may be contained in a formulation that provides the subject (patient) with an immediate release, sustained release, delayed release after administration or any other release profile known to those skilled in the art. The mode of administration and mode of administration selected for a given treatment are closely related to factors such as the mental state and physical condition of the subject (patient) as well as the amount of treatment of the compound or composition that is desirable and effective for a given treatment.

The pharmaceutical compositions of the present invention may be manufactured, packaged, or sold in bulk, as a single unit dose, as a plurality of single unit doses, or as multiple dosage forms. As used herein, a "unit dose" is a discontinuous amount of a pharmaceutical composition comprising a predetermined amount of the active ingredient. The amount of active ingredient in each unit dose generally corresponds to the total amount of active ingredient to be administered or a convenient fraction of the total dose, such as, for example, one-half or one-third of the dose. Formulations of the pharmaceutical compositions of the invention suitable for oral administration may be in the form of discrete solid dosage units. Each solid dosage unit contains a predetermined amount of the active ingredient, e.g., a unit dose or fraction thereof. As used herein, an "oily" liquid is one comprising a carbon or silicon-based liquid having a polarity less than that of water. In the above pharmaceutical dosage form, the active agent is preferably used in combination with one or more pharmaceutically acceptable carrier (s) and optionally other therapeutic ingredients. The carrier (s) should be pharmaceutically acceptable in terms of compatibility with the other ingredients of the formulation, and should not be unduly harmful to its receptor. The compositions of the present invention may be presented in unit dosage form, wherein each unit dose, such as a teaspoon, tablet, capsule, solution or suppository, may contain a predetermined amount of the active drug or prodrug alone or in combination with other pharmaceutically active agents . The term "unit dosage form" refers to suitable physical discrete units as a single administration to human and animal subjects, each unit containing a predetermined amount of the composition of the present invention alone or in an amount sufficient to produce the intended effect In combination with other active agents.

Dosage forms of the pharmaceutical compositions of the present invention may be produced by techniques known in the art and contain a therapeutically effective amount of the active compound or ingredients. Any technique known or later developed may be used in the preparation of the pharmaceutical compositions or preparations according to the invention. Generally, the preparation involves incorporating the active ingredient into a carrier or one or more other additional ingredients, and then, if necessary or appropriate, shaping the product into the desired single- or multiple-dosage unit. The powder and granule preparation according to the present invention can be produced by using a known method or a developed method. The formulations can be administered directly to the subject or can be used to produce aqueous or oily suspensions or solutions, for example by forming tablets, filling capsules, or by addition of an aqueous or oily vehicle thereto. Tablets may optionally be produced by compression or molding with one or more accessory ingredients or by wet granulation. Compressed tablets may be produced by compressing the active ingredient in a suitable device, such as a powder or granules, in a flowable form. Molded tablets may be produced by molding a mixture of the active ingredient, a pharmaceutically acceptable carrier and at least a liquid sufficient to wet the mixture in a suitable device. The tablets may not be coated or may be coated using methods known or developed in the art. Coated tablets may be formulated for delayed disintegration in the gastrointestinal tract of a subject, for example, using enteric coated tablets to provide sustained release and absorption of the active ingredient. The tablets may further comprise ingredients for providing pharmaceutically elegant and tasty preparations. Hard capsules containing the active ingredient may be produced using a physiologically degradable composition such as gelatin. Such hard capsules may contain the active ingredient. Soft gelatin capsules containing the active ingredient may be produced using a physiologically degradable composition, such as gelatin. The soft capsule comprises an active ingredient which can be mixed with water or an oil medium. Liquid preparations of the pharmaceutical compositions of the present invention suitable for administration may be prepared, packaged and sold in the form of a dry product to be reconstituted in liquid form or using water or another suitable vehicle before use. Liquid suspensions in which the active ingredient is dispersed in an aqueous or oily vehicle and liquid solutions in which the active ingredient is dissolved in an aqueous or oily vehicle can be prepared using conventional methods or developed methods. The liquid suspension of the active ingredient may be present in an aqueous or oily vehicle. Liquid solutions of the active ingredients may be present in the aqueous or oily vehicle. To produce a pharmaceutical dosage form, the active ingredient, e.g., naphthofuran, may be intimately mixed with the pharmaceutical carrier by conventional pharmaceutical compounding techniques. The carrier may take a variety of forms depending on the form of preparation required for administration. In order to produce the compositions in oral dosage form, any of the usual pharmaceutical media may be employed.

In some embodiments of the invention, the article of manufacture comprises a container containing a therapeutically effective amount of a pharmaceutical composition comprising a compound according to Formula < RTI ID = 0.0 > I. < / RTI > The container may comprise a pharmaceutically acceptable excipient. The container may contain instructions on a printed label. For example, the printed label may indicate the dose and frequency with which the pharmaceutical composition is to be administered and whether the composition should be administered with the food or within a defined time period before or after food ingestion. The composition may be contained in any suitable container capable of maintaining and quantitating dosage forms that do not substantially interact with the composition. Label instructions may be consistent with the treatment methods described herein. The label may be associated with the container by means of maintaining the physical proximity of the two. By way of non-limiting example, containers and labels may be housed in a package, e.g., a box or a plastic shrink wrap, or may include instructions to be bonded to a container using an adhesive or other bonding or holding means that does not obscure the labeling instructions, And may be combined with other matters.

In some embodiments of the invention, the pharmaceutical composition comprises (a) a therapeutically effective amount of an active ingredient that is a compound of the invention, for example, a compound according to Formula I, (b) a compound having a hydrophilic- lipophilic balance (HLB) (C) a polyoxyl glyceride having an HLB of less than 10. More preferably, the pharmaceutical composition further comprises (d) a surfactant.

Preferable examples of the polyoxyl glyceride having an HLB of more than 10 include HLB of 10 to 17, and more preferably 12 to 15 of HLB. Further preferred examples are those which are solid or semi-solid at 25 占 폚, preferably those having a melting point of more than 30 占 폚, more preferably one of the melting points is 33 to 64 占 폚, more preferably one of the melting points is 40 -55 < 0 > C. Specific examples include lauroylpolyoxyl glycerides, more specifically lauroylpolyoxyl-32 glycerides, such as Geluluxe 44/14 and stearoylpolyoxyl glycerides, and more specifically stearoylpolysiloxyl -32 glycerides, such as Gelusil ™ 50/13, are preferred. A more preferred example is lauroylpolyoxylglyceride, more specifically lauroylpolyoxyl-32 glyceride, such as Geluluxe 44/14.

Preferable examples of the polyoxyl glyceride having an HLB of less than 10 include those having an HLB of 2 to 8, and more preferably those having an HLB of 3 to 7. [ Specific examples include linoleoyl polyoxyl glycerides, such as Labrafil (TM) M2125CS, Leoyl polyoxyl glycerides such as Labrafil (TM) M1944CS and Lauroyl polyoxyl-6 glycerides such as Labrafil (TM) M2130CS. More specific examples include linoleoyl polyoxyl glycerides and oleoyl polyoxy glycerides.

Examples of surfactants include sodium lauryl sulfate (SLS) or sodium dodecyl sulfate (SDS), polyoxyethylene sorbitan fatty acid esters (polysorbate, preferably polyoxyethylene sorbitan monooleate (TWEEN 80 ™ ) Or polyoxyethylene sorbitan monolaurate (TWEEN 20 (TM)), a specific lipid such as phosphatidylcholine, for example, dimyristoylphosphatidylcholine (DMPC). Surfactants include amphoteric compounds, The preferred surfactant is sodium lauryl sulfate (SLS) or sodium dodecyl sulfate (SDS).

The active ingredient may be included in the range of 5% to 50% by weight of the formulation. Surfactants may be included in the range of 0.05% to 5% by weight of the formulation. The polyoxyl glyceride having an HLB of more than 10 may be contained in the range of 5% to 80% with respect to the weight of the preparation. A polyoxyl glyceride having an HLB of less than 10 may be included in the range of 5% to 80% with respect to the weight of the preparation. The ratio between the polyoxyl glycerides with an HLB greater than 10 and the polyoxyl glycerides with an HLB less than 10 is from about 90/10 to about 10/90. Preferably, the ratio is from about 80/20 to about 20/80, more preferably from about 40/60 to about 80/20. The composition may comprise about 27.18% active ingredient by weight, about 0.27% surfactant, about 14.51% polyoxyl glyceride having an HLB greater than 10, and about 58.04% HLB less than 10 polyoxyl glycerides. A 125 mg capsule embodiment may consist of 125 mg of active ingredient, about 1.2 mg of a surfactant, about 66.8 mg of a polyoxyl glyceride with an HLB above 10 and about 267 mg of a polyoxyl glyceride with an HLB of less than 10. An 80 mg capsule embodiment may consist of about 80 mg of active ingredient, about 0.8 mg of a surfactant, about 42.7 mg of polyoxyl glyceride with an HLB above 10, and about 170.9 mg of a polyoxyl glyceride with an HLB of less than 10. Embodiments of the present invention include any article of manufacture in which any of the above pharmaceutical compositions are encapsulated, e. G., In a LIcap capsule. It is preferred that the capsules have size 1 or smaller, e.g., size 2.

Process for the preparation of pharmaceutical preparations having a selected particle size distribution and method

Milling process

In the process according to the invention, the particle size of the active ingredient or the compound according to formula I can be reduced using a milling or grinding process. For example, the milling or milling process may be suitable for producing particles having a median size or larger or smaller size of 200 μm, 150 μm, 100 μm, 40 μm, 20 μm, 5 μm, 2 μm. Examples of the milling or crushing process include ball milling, roll milling, jet milling, wet milling, ultrasonic milling, crushing and combination. For example, such a process can impinge particles on a hard surface or treat the particles at high pressure, for example by squeezing particles between two surfaces to reduce particle size. For example, in jet milling, gas flow entrains particles and accelerates it at high speed. The particles then collide with other particles and walls and rupture into smaller particles. For example, in wet grinding, the particles are combined with a liquid and the resulting slurry is passed through a high shear mixer to rupture the particles. For example, in ultrasonic milling, for example, the particles in the slurry are exposed to ultrasonic radiation. Cavitation induced by ultrasonic waves can rupture particles into smaller sized particles.

It may be advantageous to warm the temperature of the particles before the particles are subjected to a milling or grinding operation. For example, the particles can be exposed to a cryogenic fluid, such as liquid nitrogen, or immersed, thereby warming the temperature to a cryogenic temperature. Such warming can make the particles more likely to crumble, and may increase the likelihood of having their size reduced in milling or grinding operations. After the milling or grinding process, a selection process, such as a sieving process, can be used to narrow the particle size range.

Crystallization process

Crystallization is the main separation and purification step for the preparation of drug particles. Crystallization can also be used to control particle size. The particle size distribution (PSD) obtained during crystallization may be affected by a combination of various mechanisms occurring during crystallization, such as nucleation, growth, agglomeration, wear, decomposition, Control of the PSD during crystallization is important in achieving the desired product properties. When the particle size can not be consistently controlled during crystallization to meet the desired requirements, it may include additional processing steps, such as dry milling. Braat, et al., Crystallization: Particle Size Control, Encyclopedia of Pharmaceutical Technology: Third Edition, Published on 02 October 2006].

How to treat cancer

The method according to the present invention for the treatment of symptoms of a person suffering from neoplasia, the treatment of symptoms of a mammalian or animal subject, the delay of progression, the prevention of the recurrence or the improvement of relapse slows the growth of neoplasia, A therapeutically effective amount of a pharmaceutical composition comprising particles of a predetermined size distribution to stop, reduce the volume of neoplasia, and / or kill cancerous neoplasia, for example a compound according to formula I, , Pure compounds, pure products and / or pure pharmaceutical compositions. Some examples of the types of neoplasms that can be treated by this method include solid tumors, malignant tumors, cancers, intractable cancers, recurrent cancers, metastatic tumors, neoplasms including cancer stem cells, Neoplasms, carcinomas, and sarcomas that have been identified. In some embodiments, the cancer that can be treated by administration of the particle of the compound according to Formula I is selected from the group consisting of esophageal cancer, gastric adenocarcinoma, glioma adenocarcinoma, chondrosarcoma, colon cancer, colon adenocarcinoma, rectal adenocarcinoma, Breast cancer, ovarian cancer, head and neck cancer, melanoma, gastric adenocarcinoma and adrenocortical carcinoma. The STAT3 pathway may be involved in these cancers. The CSC pathway may be involved in these cancers.

In an embodiment of the present invention, a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, is administered to a patient or subject diagnosed with a cancer, wherein the cancer is a gastro- Cancer, esophageal cancer, or gastroesophageal adenocarcinoma. Optionally, an anti-mitotic agent such as paclitaxel is administered as a second / concomitant agent for combination therapy. In one aspect, a compound of the invention is administered to a subject in a total daily dosage within the range of from about 160 mg to about 1,000 mg, preferably from about 4 hours to about 16 hours between administration of the compound, Administered as BID within a range of about 12 hours. The optional co-drug paclitaxel may be administered to a subject in a total weekly dosage ranging from about 40 mg / m 2 to about 100 mg / m 2, such as about 80 mg / m 2.

Cancer stem cell

In recent years, a novel model of tumorigenesis has been widely accepted, where only a small fraction of the entire tumor mass is hypothesized to be due to tumorigenic activity in tumors, whereas conventional or clonal genetic models have shown that mutated tumor cells Was considered to contribute equally to the above-mentioned tumorigenic activity. This small fraction of tumorigenic cells by the novel model transforms the cells into stem-cell-like amounts, which are referred to as "cancer stem cells" (CSC). Bonnet and Dick first demonstrated the presence of CSC in acute myelogenous leukemia (AML) in vivo in the 1990s. Their data indicate that only a small subset of human AML cells have the ability to transduce AML upon implantation into immunodeficient mice and that other AML cells can not induce leukemia. Subsequently, these CSCs were found to have the same cellular markers CD34 ⁺ / CD38 ^- as primitive hematopoietic stem cells. Bonnet, D., Normal and leukemic stem cell. Br J Haematol , 2005. 130 (4): p. 469-79]. Since then, researchers have found CSC to be crucial in various types of tumors, including brain, breast, skin, prostate, and colon cancer.

The CSC model of tumorigenesis explains why hundreds of thousands or hundreds of thousands of tumor cells must be injected into experimental animals to establish tumor grafts. In human AML, the frequency of these cells is less than 1 in 10,000. Bonnet, D. and JE Dick, Human acute myeloid leukemia is organized as a hierarchy originating from a primitive hematopoietic cell. Nat Med , 1997. 3 (7): p. 730-7]. Although rare in a given tumor cell population, there is growing evidence that the cells are present in almost all tumor types. However, since the cancer cell line is selected from a sub-population of cancer cells specifically modified to grow in the tissue culture fluid, the biological and functional properties of the cancer cell line may undergo drastic changes. Therefore, not all cancer cell lines contain CSC.

Cancer stem cells share many similar traits with normal stem cells. For example, CSC has the ability to cause additional tumorigenic stem cells at a slower rate than other dividing tumor cells, as opposed to self-renewing capacity, a so-called limited number of divisions. CSCs also have the ability to differentiate into multiple cell types, which not only contain multiple cell types that are unique to a host organism, but also describe histological evidence that heterogeneity is normally retained in tumor metastasis . CSC has been shown to be fundamentally attributable to tumorigenesis, cancer metastasis and cancer recurrence. CSCs are also referred to as tumor-initiated cells, cancer stem-like cells, stem-like cancer cells, high tumorigenic cells, tumor stem cells, solid tumor stem cells or highly malignant cells.

The presence of cancer stem cells has a fundamental implication for future cancer treatment and therapy. These events are identified by disease identification, selective drug targeting, prevention of cancer metastasis and recurrence, and the establishment of new strategies to combat cancer.

The efficacy of current cancer therapies is often measured by the magnitude of tumor shrinkage, the amount of tumor mass that is killed, in the early stages of the test. Since CSCs form a very small proportion of tumors and have significantly different biological characteristics than their more differentiated descendants, the measurement of tumor mass is not necessarily selectable for drugs that act specifically on stem cells. In fact, cancer stem cells exhibit resistance to radiation therapy (XRT) and also refractory to chemotherapeutic agents and targeted drugs. Hambardzumyan, D., M. Squatrito and EC Holland, Radiation resistance and stem-like cells in brain tumors. Cancer Cell , 2006. 10 (6): p. 454-6; Baumann, M., M. Krause and R. Hill, Exploring the role of cancer stem cells in radioresistance. Nat Rev Cancer , 2008. 8 (7): p. 545-54; Ailles, LE and IL Weissman, Cancer stem cells in solid tumors. Curr Opin Biotechnol , 2007. 18 (5): p. 460-6]. Normal somatic stem cells are naturally resistant to chemotherapeutic agents, which have a variety of pumps (e.g., MDR) that pump drugs and DNA repair proteins. In addition, they also have a slow cell alternation rate, while chemotherapeutic agents rapidly target cells that are replicating. Cancer stem cells, which are mutated counterparts of normal stem cells, may also have a similar mechanism to withstand drug therapy and radiation therapy. In other words, conventional chemotherapy and radiation therapy differentiate or kill cells during differentiation, forming a bulk of tumors that are unable to produce new high tumorigenic cancer stem cells. On the other hand, the population of cancer stem cells that produce differentiated or differentiating cells remains intact and causes a recurrence of the disease. An additional risk for conventional chemotherapy is the possibility that chemotherapy treatments will leave only chemotherapy-resistant cancer stem cells, and the subsequent recurrent tumors will be resistant to chemotherapy.

Since surviving cancer stem cells can cause tumor recurrence and cause recurrence, chemotherapy should include a strategy for CSC (see Figure 18 of WO 2011/116398 and WO 2011/116399). This is similar to rooting out to prevent the dandelion from regrowing, even though cutting off the clumps of weed soil. [Jones, RJ, WH Matsui and BD Smith, Cancer stem cells: are we missing the target? J Natl Cancer Inst , 2004. 96 (8): p. 583-5]. Cancer stem cells can be selectively targeted to treat patients with aggressive, non-resectable tumors and intractable or recurrent cancers, as well as prevent tumor metastasis and recurrence. The development of specific therapies targeting cancer stem cells can improve the quality of life and survival of cancer patients, especially in patients with metastatic cancers. The key to releasing these undeveloped possibilities is identification and authentication of pathways that are selectively critical for cancer stem cell self-renewal and survival. Unfortunately, a number of pathways underlying tumorigenesis in cancer or self-renewal in embryonic and adult stem cells have been identified in the past, but very few pathways have been identified and validated for cancer stem cell self-renewal and survival .

In addition, a number of studies have been conducted on identification and isolation of cancer stem cells. The predominant method used is based on the ability of the CSC to spill the drug or the expression of surface markers associated with cancer stem cells.

For example, since CSCs are resistant to a number of chemotherapeutic agents, CSC can be used as a drug efflux pump, such as ABCG2 (BCRP-1) (Ho, MM, et al., Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells cancer Res, 2007. 67 (10):... p 4827-33; Wang, J., et al, Identification of cancer stem cell-like side population cells in human nasopharyngeal carcinoama cell line . cancer Res, 2007. 67 (8 ):... p 3716-24; Haraguchi, N., et al, Characterization of a side population of cancer cells from human gastrointestinal system Stem cells, 2006. 24 (3): p .. 506-13; Doyle, LA and DD Ross, Multidrug resistance mediated by the breast cancer resistance protein BCRP (ABCG2) Oncogene, 2003. 22 (47):.. p 7340-58; Alvi, AJ, et al, Functional . and molecular characterisation of mammary side population cells Breast Cancer Res, 2003. 5 (1):.. p R1-8) and other ATP-binding cassette (ABC) and the members (Frank, NY, et al, ABCB5-mediated doxorubicin transport and Cancer Res , 2005. 65 (10): 4320-33; Schatton, T., et al., Identification of cells initiating human melanomas. Nature , 2008 451 (7176): p. 345-9) is over-expressed almost everywhere. Thus, an early-used side-group (SP) technique for enriching hematopoietic and leukemic stem cells has been used to identify and isolate CSCs (Kondo, T., Setoguchi and T. Taga, Persistence of a small subpopulation of cancer stem-like cells in the C6 glioma cell line. Proc Natl Acad Sci USA , 2004. 101 (3): p. 781-6). This first-described technique, by Goodell et al., Utilizes fractional ABC transporter-dependent efflux of fluorescent dyes, such as Hoechst 33342, to compartmentalize and separate dense population of cells among CSCs (Doyle, LA and DD Ross, Multidrug resistance mediated by the breast cancer resistance protein BCRP (ABCG2) Oncogene, 2003. 22 (47):... p 7340-58; Goodell, MA, et al, Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo. J Exp Med. , 1996. 183 (4): p. 1797-806). Specifically, SP is revealed by blocking the drug efflux using verapamil at a point where the dye can no longer pump the SP.

Researchers have also focused on finding specific markers that identify cancer stem cells from the bulk of tumors. Most commonly expressed surface markers by cancer stem cells include CD44, CD133 and CD166. (Collins, AT, et al., Prospective identification of tumorigenic prostate cancer stem cells. Cancer Res , 2005. 65 (23): p 10946-51; Li, C., et al., Identification of pancreatic cancer stem cells. cancer Res, 2007. 67 (3) :.... p 1030-7; Ma, S., et al, Identification and characterization of tumorigenic liver cancer stem / progenitor cells Gastroenterology, 2007. 132 (7): p 2542- 56, Prince, ME, et al., Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma., 2007. 104 (3): 973-8; Ricci-Vitiani, L. Identification and expansion of human colon-cancer-initiating cells. Natur , 2007. 445 (7123): p. 111-5; Singh, SK, et al., Identification of cancer stem cells in human brain tumors . Cancer Res, 2003. 63 (18 ):... p 5821-8; Dalerba, P., et al, Phenotypic characterization of human colorectal stem cells Proc Natl Acad Sci USA , 2007. 104 (24): p. 10158-63). Tumor cell classification based primarily on differential expression of these surface marker (s) is considered to be the major high tumorigenic CSC described to date. Therefore, these surface markers are evidenced in the identification and isolation of cancer stem cells from cancer cell lines and from bulk of tumor tissue.

Recent studies have revealed the presence of cancer stem cells (CSCs) with the exclusive ability to regenerate tumors. These CSCs are present in nearly all tumor types and are functionally related to persistent malignant growth, cancer metastasis, recurrence and cancer drug resistance. CSC and its more distinct descendants appear to have significantly different biological characteristics. Conventional cancer drug screening relies on the measurement of the amount of tumor mass, so that it does not necessarily have to be selected for drugs that specifically work against CSC. In fact, CSCs are resistant to standard chemotherapy and radiotherapy and have proved to be enriched after standard chemotherapy, resulting in cancer refractory and recurrence. Examples of the method for separating these cells include identification by their efflux capacity of their chromosome 33342, confirmation by surface markers expressing these cells, such as CD133, CD44, CD166, etc., and their enrichment by tumorigenic properties. It is not limited. Increasing evidence linking cancer stem cells to tumorigenesis addresses a tremendous therapeutic opportunity to target cancer stem cells.

The data provided herein, along with recent breakthroughs in the CSC studies, suggests that the present invention is directed to a method involving the inhibition of both CSC and heterogeneous cancer cells in a number of ways involving inhibition of CSC and, And a method for treating the disease. The present invention also provides related methods (e.g., manufacturing and drug candidate screening), materials, compositions and kits. The method can prevent CSC from self-regenerating such that it can not divide into tumorigenic CSC cells so that its number can no longer be supplemented. Alternatively, the method can induce cancer death in CSC or in both CSC and heterogeneous cancer cells.

Such a method can be used to treat cancer of a subject. Cancers that are excellent candidates for the treatment include esophageal cancer, gastric adenocarcinoma, glioma adenocarcinoma, chondrosarcoma, colon cancer, colon adenocarcinoma, rectal adenocarcinoma, colon adenocarcinoma, breast cancer, ovarian cancer, head and neck cancer, melanoma, Cancer (s) selected from the group consisting of adrenocortical carcinomas.

In addition, since CSC has been demonstrated to be fundamentally attributable to tumorigenesis, cancer metastasis and cancer recurrence, any of the methods of the present invention for inhibiting CSC or both CSC and heterogeneous cancer cells can be used to treat metastatic, May be performed to treat cancer that is refractory to therapy or recurrent in a subject after initial treatment.

In some embodiments, the cancer stem cell inhibitor according to the present invention comprises a compound of formula 1, a compound 1, a polymorph of compound 1, an X-ray diffraction pattern substantially similar to that depicted in Figure 1 of WO 2011/116398 and WO 2011/116399 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, which is substantially similar to that shown in Figure 2 of WO 2011/116398 and WO 2011/116399 A polymorph of 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione characterized by an X-ray diffraction pattern, a peak at at least about 10.2 degrees 2 theta, A peak at least at about 14.1 degrees two theta, at least at about 14.5 degrees two theta, at least at about 17.3 degrees two theta, at least at about 22.2 degrees two theta, and at least about 28.1 degrees two theta, Acetyl-4H, 9H75 naphtho [2,3-b] furan-2-one, characterized by an X-ray diffraction pattern comprising at least two peaks from any combination of & 2-acetyl-4H, 9H-naphtho [2,3-dione], which is characterized by an X-ray diffraction pattern substantially similar to that shown in Figure 3 of WO 2011/116398 and WO 2011/116399, b] furan-4,9-dione, a peak at at least about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 12.3 degrees two theta, a peak at least about 15 degrees two theta, A peak at about 23 ° 2?, A peak at at least about 23.3 ° 2 ?, a peak at at least about 24.6 ° 2 ?, and a peak at least about 28.4 ° 2? 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, 2- (1-hydroxyethyl) -naphtho [2,3- b ] Furan-4,9-dione, 2-acetyl-7-chloro-naphtho [2,3- b] b] furan-4,9-dione, 2-acetylnaphtho [2,3- b] furan-4,9-dione, 2-ethyl-naphtho [2,3- b] , Vinyl] ester, phosphoric acid 1- (4-fluoro-4-fluoro-phenyl) , 9-dioxo-3a, 4,9,9a-tetrahydro-naphtho [2,3-b] furan-2-yl) -vinyl ester dimethyl ester, its enantiomers, diastereoisomers, tautomers and salts Or polymorphs of solvates; Characterized by an X-ray diffraction pattern substantially similar to that depicted in Figure 1 of the compound of Formula 1, the polymorph of Compound 1, the polymorph of Compound 1, WO 2011/116398 and WO 2011/116399, 2-acetyl-4H, 9H -Naphtho [2,3-b] furan-4,9-dione, 2-acetyl, which is characterized by an X-ray diffraction pattern substantially similar to that shown in Figure 2 of WO 2011/116398 and WO 2011/116399 -4H, 9H-naphtho [2,3-b] furan-4,9-dione, a peak at at least about 10.2 degrees two theta, a peak at least about 11.9 degrees two theta, Peak, at least about 14.5 DEG 2 &thetas; at least about 17.3 DEG 2 DEG peak, at least about 22.2 DEG theta peak, at least about 28.1 DEG theta peak, and any combination thereof. 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, which is characterized by an X-ray diffraction pattern, A polymorph of 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione characterized by an X-ray diffraction pattern substantially similar to that specified, a peak at at least about 7.5 degrees two theta, A peak at least about 9.9 degrees two theta, a peak at least about 12.3 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 23 degrees two theta, a peak at least about 23.3 degrees two theta, Acetyl-4H, 9H-naphtho [2,3-d] pyrimidine characterized by an X-ray diffraction pattern comprising two or more peaks from a peak at at least about 28.4 DEG 2 & b] furan-4,9-dione, 2- (1-hydroxyethyl) -naphtho [2,3- b] furan-4,9-dione, 2- acetyl- 2,3-b] furan-4,9-dione, 2-acetylnaphtho [2,3-b] Furan-4,9-dione, 2-ethyl-naphtho [2,3-b] furan-4,9-dione, mono - [1- (4,9-dioxo-3a, - Tetrahydro-Naph Yl] -vinyl] ester, phosphoric acid 1- (4,9-dioxo-3a, 4,9,9a-tetrahydro-naphtho [2,3- b] Furan-2-yl) -vinyl ester dimethyl ester, its enantiomers, diastereoisomers, tautomers and salts or solvates thereof; Or a substantially pure form of a compound of formula 1, a polymorph of compound 1, a compound 1, a 2-acetyl-pyrimidine compound characterized by an X-ray diffraction pattern substantially similar to that specified in Figure 1 of WO 2011/116398 and WO 2011/116399, 4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern substantially similar to that shown in Figure 2 of WO 2011/116398 and WO 2011/116399 A polymorph of a 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, a peak at at least about 10.2 degrees two theta, a peak at least about 11.9 degrees two theta, Peaks at 2?, Peaks at at least about 14.5 2?, Peaks at at least about 17.3 2?, Peaks at at least about 22.2 2? And peaks at at least about 28.1 2? Acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione characterized by an X-ray diffraction pattern comprising at least one 8 and a polymorph of 2-acetyl-4H, 9H-naphtho [2,3-b] furan-4,9-dione, characterized by an X-ray diffraction pattern substantially similar to that specified in Figure 3 of WO 2011/116399 A peak at least at about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 12.3 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 23 degrees two theta, Characterized by an X-ray diffraction pattern comprising at least two peaks from a peak at 2 &thetas; a peak at least about 24.6 DEG 2 &thetas; and a peak at least about 28.4 DEG 2 theta, , 9H-naphtho [2,3-b] furan-4,9-dione, 2- (1-hydroxyethyl) 2,3-b] furan-4,9-dione, 2-acetyl-7-fluoro-naphtho [ Ethylnaphtho [2,3-b] furan-4,9-dione, mono - [1- (4,9-di Oxo-3a, 4,9 , 9a-tetrahydronaphtho [2,3- b] furan-2-yl) -vinyl] ester, phosphate 1- (4,9-dioxo-3a, 4,9,9a-tetrahydronaphtho [ , 3-b] furan-2-yl) -vinyl ester dimethyl ester, its enantiomers, diastereoisomers, tautomers and salts or solvates; 2,3-b] furan-4,9-dione, 2-acetyl-7-chloro-naphtho [ 2,3-b] furan-4,9-dione, 2-acetyl-7-fluoronaphtho [2,3- -Naphtho [2,3-b] furan-4,9-dione, phosphoric acid mono- [l- (4,9-dioxo-3a, 4,9,9a-tetrahydro- b] furan-2-yl) -vinyl] ester, phosphoric acid 1- (4,9-dioxo-3a, 4,9,9a-tetrahydro- -Vinyl ester dimethyl ester, its enantiomers, diastereoisomers, tautomers and salts or solvates (also referred to herein as "compounds of the invention ").

The present invention provides a method for identifying a drug candidate capable of inhibiting cancer stem cells. In some embodiments, the drug candidate can induce apoptosis in CSC or at least inhibit its self-renewal. In a further embodiment, the drug candidate can induce apoptosis in CSC, at least inhibit its self-renewal, and induce apoptosis in homogeneous cancer cells. The various steps in the pathway can be targeted to screen for drug candidates.

Thus, in another embodiment, the compounds of the present invention can be used to formulate pharmaceutical compositions for treating or preventing a disease or condition. In some embodiments, the cancer is selected from the group consisting of esophageal cancer, gastric adenocarcinoma, glioma adenocarcinoma, chondrosarcoma, colon cancer, colon adenocarcinoma, rectal adenocarcinoma, colon adenocarcinoma, breast cancer, ovarian cancer, head and neck cancer, melanoma, adenocarcinoma and adrenocortical carcinoma ≪ / RTI >

Thus, in one embodiment, the present invention provides a method of inhibiting cancer stem cells in which an effective amount of a compound of the invention is administered to a cell. Cancers known to have CSCs are excellent candidates for the treatment, including but not limited to: esophageal cancer, gastric adenocarcinoma, glioma adenocarcinoma, chondrosarcoma, colon cancer, colon adenocarcinoma, rectal adenocarcinoma, , Carcinoma (s) selected from the group consisting of head and neck cancer, melanoma, gastric adenocarcinoma and adrenocortical carcinoma.

In addition, since CSC has been demonstrated to contribute basically to tumorigenesis, cancer metastasis and cancer recurrence, any method of the present invention for inhibiting CSC is refractory to metastatic, chemotherapy, or radiation therapy, May be performed to treat recurrent cancer in a subject.

In some embodiments of the invention, the cancer being treated is selected from the group consisting of esophageal cancer, gastric adenocarcinoma, glioma adenocarcinoma, chondrosarcoma, colon cancer, colon adenocarcinoma, rectal adenocarcinoma, colon adenocarcinoma, breast cancer, ovarian cancer, head and neck cancer, Adenocarcinoma and adrenocortical carcinoma. Cancer may be associated with dysfunction of the STAT3, NANOK, and / or P-catenin pathways.

In one embodiment, the invention provides a method of treating cancer in a subject, comprising administering to the subject a therapeutically effective amount of a pharmaceutical composition comprising a compound of the invention. Cancer can be metastatic, intractable or recurrent. The subject may be a mammal, for example a human.

Treatment of a subject suffering from a neoplasm (patient) by, for example, administration of a particle of a compound according to formula I may be indicated for the following conditions. The neoplasm may be refractory to treatment with chemotherapy, radiotherapy or hormone therapy. Neoplasms may not be treated by surgical resection. The neoplasm may recur in the subject (patient). Cancer stem cells are involved in the recurrence of neoplasms; The death of cancer stem cells or the inhibition of his self-renewal by the method by name can prevent the neoplasm from regenerating itself. Treatment with the administration of the particles of naphthofuran can slow or halt the volume growth of the neoplasm or can induce the death of neoplastic cells, inhibit the growth and / or division of neoplastic cells, and / It is possible to selectively kill neoplastic cells. For example, the treatment according to the present invention may cause apoptosis of cells of neoplasia. For example, treatment can act to inhibit the STAT3, NANOK and / or &bgr; -catenin pathways of neoplastic cells.

For example, the treatment of administering the particles of the compound of the present invention to a subject suffering from a neoplasm (patient) can be used as an adjuvant therapy to prevent recurrence of neoplasia or to perform surgical resection.

For example, a pharmaceutical composition comprising particles of a compound of the present invention is a convenient form of treatment and can be administered orally. For example, the pharmaceutical composition may be orally administered more than four times a day. Alternatively, the pharmaceutical composition may be administered intravenously or intraperitoneally.

Presumed Biomarker  Used patient screening

Both phosphorylated STAT3 (p-STAT3) positivity and beta -catenin expression in the cellular nuclei, either individually or in combination, serve as predictive biomarkers for the greater potential of therapeutic efficacy with the compounds of the invention Based on the findings (see Examples 9 and 10), the present invention provides a method for screening a patient for recommendation of cancer therapy involving the compounds of the present invention. The data herein show a direct correlation between the level of p-STAT3 in tumor tissue before treatment and the survival chances of using the compounds of the invention or the success of treatment. In other words, the higher the level of p-STAT3 found in pre-treatment cancer patients at least in patients with colorectal cancer (CRC) in a single treatment using the compounds of the invention and related compositions, the higher the overall survival rate (OS) ). Thus, the present invention provides a screening method for potentially cancer patients for the treatment or treatment of cancer in a selected patient population, which method comprises the steps of administering a biological sample from a patient candidate diagnosed with cancer (for example, colorectal adenocarcinoma) Measuring the level of phosphorylated STAT3 (p-STAT3) in the tumor tissue before treatment); Confirming that the p-STAT3 level of the patient candidate is higher than a reference level; Comprising administering to a patient candidate a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof. The reference level may be different for different demographic sectors and may be determined by one of ordinary skill in the art through routine experimentation.

Similarly, since the data herein show a direct correlation between the level of expression or localization of [beta] -catenin, the oncogene is closely related to STAT3 in the nucleus and the likelihood of survival or therapeutic success using the compounds of the invention . In other words, the higher the level of β-catenin expression in the cancer cell nucleus, the higher the overall survival rate (OS), as opposed to the cell membrane in cancer patients, at least in CRC patients (FIG. 4B). Accordingly, the present invention provides a method for detecting cancer, comprising the steps of: detecting a site of? -Catenin expression in a biological sample (for example, tumor tissue before treatment) obtained from a patient candidate diagnosed with cancer; Confirming that significant beta -catenin expression is detected in the nuclei in the sample from the patient candidate; And administering to a patient candidate a therapeutically effective amount of a compound of the invention, or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, for the treatment or treatment of cancer in a selected patient population A method of screening cancer patients. Levels at which nuclear expression for < RTI ID = 0.0 > ss-catenin < / RTI > is considered clinically significant may differ for different demographic sectors and can be determined by one of ordinary skill in the art.

The invention provides a kit for detecting, or otherwise improving, e.g., stratifying, a population of patients suitable for therapeutic administration of a compound of this disclosure by detecting the level of expression of one or more biomarkers associated with cancer stem cells, and / Provide a kit. In methods and / or kits of the present disclosure, the level of expression of one or more cancer stem cell markers is detected in a sample from a patient or a patient, and wherein the patient or sample has one or more cancer stem cell markers If the patient has an increased level, the patient is administered a therapeutic dose of the compound of the disclosure. In some embodiments of these methods, the method is an in vivo method. In some embodiments of these methods, the method is an in situ method. In some embodiments of these methods, the method is an in vitro method. In some embodiments of these methods, the method is an in vitro method.

Understanding the clinical relevance of cancer stem cell markers and identifying its predictive response to the compounds of the present invention is used herein to support clinical development by selecting patients who are most likely to derive the clinical benefit. In some embodiments, the methods provided herein employ expression of one or more known cancer stem cell marker (s), such as p-STAT3 and / or other cancer stem cell-related proteins, such as beta -catenin and nanor . All of these proteins can be readily detected using any of the various techniques known in the art. In some embodiments, cancer stem cell markers are detected by immunohistochemistry using antibodies. Patient responses to the compounds of the present invention were analyzed based on biomarker status using archival tissue samples collected from the Phase I test using compounds of the invention. Analysis of treated CRC patients using the compounds of the present invention demonstrated a trend of increased survival for patients with high levels of p-STAT3 or NARK compared to patients with low or negative levels of p-STAT3 or NARAC Respectively. Significant improvements in survival were detected in patients with nuclear β-catenin localization compared to patients with localized β-catenin on the cell membrane. HR = 0.043, p value < 0.001. As additional evidence, in vitro experiments in screening panels of cancer cells, cell lines with nuclear beta -catenin exhibit lower IC ₅₀ for the compounds of the present invention. In addition, inhibition of STAT3 by the compounds of the present invention reduces beta -catenin protein levels in both cancer cell lines and human CRC xenografted mouse models. Thus, STAT3 activation is involved in the regulation of nuclear beta -catenin. In addition, the beta -catenin state is a biomarker for predicting the responsiveness of a CRC patient to a compound of the invention.

In various embodiments of the methods of treatment, the cancer is selected from the group consisting of esophageal cancer, gastric adenocarcinoma, glioma adenocarcinoma, chondrosarcoma, colon cancer, colon adenocarcinoma, rectal adenocarcinoma, colon adenocarcinoma, breast cancer, ovarian cancer, head and neck cancer, melanoma, And adrenocortical carcinoma. Cancer can be refractory, recurrent or metastatic.

Drug therapy, dosage and interval

In a method according to the present invention, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or a purified form of a compound of the present invention is about 20 mg to about 2000 mg, about 100 mg to about 1,500 mg, From about 160 mg to about 1400 mg, or from about 180 mg to about 1200 mg. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph, and / or a purified form of a compound of the present invention is in the range of from about 200 mg to about 1500 mg, or from about 360 mg to about 1200 mg, Dose. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or a purified form of a compound of the invention is a total daily dose within the range of from about 400 mg to about 1,000 mg. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or a purified form of a compound of the invention is a total daily dose of about 1,000 mg.

The interval between each administration can be varied depending on the requirements, such as the pharmacokinetics of the composition, the drug metabolism of the fluid or food ingestion or ingestion, the tolerability and other drug-taking fidelity factors (e.g., simplicity) Or can be kept constant. Preferred intervals maintain an effective level of the pharmaceutical composition in the body while causing minimal side effects. In some embodiments, the interval between each administration is within the range of about 4 hours to about 24 hours. In some embodiments, the interval between each administration is in the range of about 8 hours to about 14 hours. In some embodiments, the interval between each administration is in the range of about 10 hours to about 13 hours, or about 12 hours. Thus, in these embodiments, the compound is administered to the subject, for example, about two times a day, on average, over the period of therapy.

In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph, and / or a purified form of a compound of the present invention is a total daily dose within the range of about 160 mg to about 960 mg or about 1,000 mg . In some embodiments, the therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the invention is about 160 mg, about 320 mg, about 640 mg, about 800 mg, and about 960 mg Lt; RTI ID = 0.0 &gt; 1 &lt; / RTI &gt; daily dose. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or a purified form of a compound of the invention is administered to a subject at a total daily dose of about 960 mg.

In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the invention is administered BID. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph, and / or a purified form of a compound of the invention is administered to a subject in a dosage ranging from about 80 mg BID to about 480 mg BID. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph, and / or a purified form of a compound of the invention is administered to a subject at a dose of about 80 mg BID, about 160 mg BID, about 320 mg BID, Mg BID and about 480 mg BID. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the invention is administered to a subject at a dose of about 480 mg BID.

In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the invention is administered BID, and the time between administrations is from about 4 hours to about 4 hours It is within about 16 hours. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph, and / or a purified form of a compound of the invention is administered BID and the time between administration of the compound is greater than 4 hours, greater than 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours and / or 16 hours. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or a purified form of a compound of the invention is administered to a subject at a dose within the range of about 80 mg BID to about 480 mg BID, The time between administration of the compound is within the range of about 4 hours between administrations to about 16 hours between administrations. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph, and / or a purified form of a compound of the invention is administered BID and the time between administration of the compound is greater than 4 hours, greater than 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours and / or 16 hours. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph, and / or a purified form of a compound of the invention is administered to a subject in an amount of about 80 mg, about 160 mg, about 320 mg BID, about 400 mg BID And about 480 mg BID, the time between administration of the compound being in the range of about 4 hours between administrations to about 16 hours between administrations. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph, and / or a purified form of a compound of the invention is administered BID and the time between administration of the compound is greater than 4 hours, greater than 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours and / or 16 hours. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph, and / or a purified form of a compound of the invention is administered to a subject at a dose of about 480 mg BID, the time between administration of the compound Is within about 4 hours to about 16 hours between administrations. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the invention is administered to a subject at a dose of about 80 mg BID, the time between administration of the compound Is within about 4 hours to about 16 hours between administrations. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the invention is administered to a subject at a dose of about 400 mg BID, the time between administration of the compound Is within about 4 hours to about 16 hours between administrations. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the invention is administered to a subject at a dose of about 320 mg BID, the time between administration of the compound Is within about 4 hours to about 16 hours between administrations. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph, and / or a purified form of a compound of the invention is administered BID and the time between administration of the compound is greater than 4 hours, greater than 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13 hours, 14 hours, 15 hours and / or 16 hours.

In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the invention is administered to a subject at a dose of about 480 mg BID, the time between administration of the compound Is about 12 hours between administration. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the invention is administered to a subject at a dose of about 80 mg BID, the time between administration of the compound Is about 12 hours between administration. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the invention is administered to a subject at a dose of about 400 mg BID, the time between administration of the compound Is about 12 hours between administration. In some embodiments, a therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the invention is administered to a subject at a dose of about 320 mg BID, the time between administration of the compound Is about 12 hours between administration.

The compounds of the present invention or pharmaceutical compositions thereof may be administered by any route or combination thereof, for example, orally, intravenously or intraperitoneally. For example, in some embodiments, the compounds of the invention may be administered orally. In some embodiments, the compounds of the present invention are formulated with a formulation comprising lauroylpolyoxylglyceride (e.g., gelulose) and Tween 80 or a formulation comprising lauroylpolyoxylglyceride (e.g., gelulose), linoleylpolyoxyl Orally administered with a formulation comprising a glyceride (e.g., labrafil) and a surfactant such as sodium lauryl sulfate (SLS) or sodium dodecyl sulfate (SDS).

The compounds of the present invention may be administered at doses that achieve a concentration of the compound in a subject, e. G., A patient, in the range of about 0.002 μM to about 30 μM for a time of 2 hours up to 24 hours. In some embodiments, a compound of the invention is administered at about 0.2, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 4.0, At a dose that achieves a blood concentration in a subject of a compound in the range of at least about 0.2 μM to about 1 μM for a time ranging from 2 hours to 24 hours, corresponding to 8.0 μM, 9.0 μM, 10.0 μM, 15.0 μM or more &Lt; / RTI &gt; In some embodiments, a compound of the invention has a plasma concentration in a subject of a compound corresponding to at least about 1.0 μM, 1.5 μM, 2.0 μM, 3.0 μM, 5.0 μM, 10.0 μM, and 15.0 μM for greater than or equal to 2 hours and less than 24 hours &Lt; / RTI &gt; In some embodiments, the compounds of the invention may be administered at doses that achieve a plasma concentration in a subject of a compound corresponding to at least about 2.0 μM, 3.0 μM, 5.0 μM, 10.0 μM for greater than or equal to 2 hours and less than 24 hours have. In some embodiments, a compound of the invention may be administered at a dose to achieve a blood concentration in a subject of a compound corresponding to at least about 3.0 [mu] M or 5.0 [mu] M for at least 2 hours and less than 24 hours.

The compounds of the present invention may be administered within 24 hours at a dosage to achieve a concentration of the compound in a subject, e.g., a patient, in the range of at least about 0.002 μM.h to about 300 μM.h. In some embodiments, a compound of the invention is administered at about 0.2, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 4.0, 4.0, , 8.0 [mu] M, 9.0 [mu] M, 10.0 [mu] M, 15.0 [mu] M or more. In some embodiments, the compound of the invention has a plasma concentration in a subject of a compound corresponding to at least about 1.0, 1.5, 2.0, 3.0, 5.0, 10, or 15.0 [mu] M for greater than or equal to 2 hours and less than 24 hours &Lt; / RTI &gt; In some embodiments, the compounds of the invention may be administered at doses that achieve a plasma concentration in a subject of a compound corresponding to at least about 2.0 μM, 3.0 μM, 5.0 μM, 10.0 μM for greater than or equal to 2 hours and less than 24 hours have. In some embodiments, a compound of the invention may be administered at a dose to achieve a blood concentration in a subject of a compound corresponding to at least about 3.0 [mu] M or 5.0 [mu] M for at least 2 hours and less than 24 hours. In some embodiments, the compounds of the invention are administered at a dose of about 2 μM * hr, 10 μM * hr, 20 μM * hr, 30 μM * hr, 40 μM * hr, 50 μM * hr, 60 μM * hr, , 80 μM * hr, 90 μM * hr, 100 μM * hr, 125 μM * hr, 150 μM * hr, 200 μM * hr, 250 μM * hr, 300 μM * hr, 400 μM * hr and 500 μM * hr (AUC _{0-24 hours} ) below the curve within 24 hours in the corresponding subject.

If the condition of the subject (patient) is so desired, the dosage of the pharmaceutical composition may be administered as a continuous or pulsatile injection. The treatment period may be several decades, several years, months, weeks or even days as long as the benefit lasts. The range is provided as a guideline only and is optimized.

In the method according to the invention, the cells of the neoplasm are selectively killed by the administration of the pharmaceutical composition, so that the blood molar concentration of the compound is at least as long as the effective time, above the effective concentration for the first consecutive period, , Lower than harmful concentration. The molar concentration in the blood may be lower than the effective concentration after the first continuous period. The effective concentration can be a sufficiently high temperature to kill neoplastic cells, such as cancer cells. The lifespan can be long enough to kill neoplastic cells, such as cancer cells. The harmful concentration may be a concentration that damages or kills normal cells. The detoxification time may be long enough for normal cells to become damaged or killed. For example, the effective concentration may be about 0.02 μM, about 0.05 μM, about 0.1 μM, about 0.2 μM, about 0.5 μM, about 1 μM, about 3 μM, about 10 μM, or about 20 μM or more. For example, the non-hazardous concentration may be about 3 μM, about 10 μM, about 14 μM, about 30 μM, or about 100 μM or less. For example, the shelf life may be about 2 hours, about 4 hours, about 6 hours, about 12 hours, about 24 hours, or about 48 hours or more. For example, to achieve a non-hazardous exposure to normal cells, the drug concentration of Compound 1 should be substantially cleaned from the blood within about 12 hours, about 24 hours. "Substantial cleansing from blood" means a blood drug concentration reduction of at least about 50%, at least about 60%, at least about 80%, at least about 90%. For example, the effective concentration may be a concentration that exceeds the IC ₅₀ of cancer cells upon administration of the compound over a period of time. For example, the effective time may be the time to selectively inhibit or kill the cancer cells upon administration of the compound at least at an effective concentration. For example, the harmful concentration may be a concentration at which the compound exceeds the IC ₅₀ of normal cells upon administration for any period of time. For example, the harmful time may be the time at which the cancer cells as well as the normal cells are inhibited or killed when the compound is administered at an effective concentration.

Those skilled in the art will appreciate that by selecting the dosage and frequency to achieve a "selective pharmacokinetic profile" (SPP) described herein that appears to be necessary for selectively killing neoplastic cells, such as cancer cells, Lt; / RTI &gt; The above considerations of SPP can also guide the design of pharmaceutical compositions, such as particle size distribution and particle shape distribution.

In the method according to the present invention, the pharmaceutical composition may be formulated into dosage forms such as tablets, pills, capsules (hard or soft), dragees, powders, granules, suspensions, liquids, gels, cachets, troches, lozenges, syrups, , Emulsions, oil-in-water emulsions, water-in-oil emulsions or potions.

Identification of optimal particle size distribution

In the method according to the invention, the compound according to formula I for the treatment of a person suffering from neoplasia, mammal or animal, the compound 1, the polymorph of compound 1 and / or the optimal particle of substantially pure form of compound 1 The size distribution can be measured as follows. One or more sets of particles comprising the compound. In preparing a set of particles, for example, the particle size of a sample of a solid compound can be determined by, for example, dissolving the compound, spraying the solution, dissolving the compound and ultrasonifying the solution, ball milling the solid compound, May be reduced by roll milling, pulverizing the solid compound and / or sieving the solid compound. The particle size distribution of one or more sets of particles can be determined by a method or combination of methods known to those skilled in the art. For example, the particle size distribution can be measured using techniques such as sieve analysis, optical microscopy, electron microscopy, electrical resistivity, settling time, laser diffraction and / or acoustic spectroscopy, or any other technique or combination of techniques . One or more sets of particles may be administered to the neoplastic and normal cells for a predetermined time at a predetermined concentration. The effect of particles on metabolism, cleavage and / or other indicators of vigor of neoplastic and normal cells can be observed. The observed effect of particles on neoplastic cells can be used to assign an efficiency rating to each set of particles. For example, a set of particles that inhibit metabolism and / or division of neoplastic cells, damage or kill neoplastic cells, or otherwise exhibit high antineoplastic activity, can be assigned a high efficiency rating. The observed effect of particles on normal cells can be used to assign a toxicity assessment to each set of particles. For example, a set of particles that inhibit the metabolism and / or division of normal cells, or that impair or kill normal cells, or that exhibit a low tolerance of the particle set if normal cells are not, can assign a high toxicity assessment .

For example, a set of particles can be administered in vitro to neoplastic cells and normal cells. For example, an efficiency assessment may correspond to, or proportionally or monotonically increase, the action of an IC ₅₀ of a neoplastic cell. For example, the toxicity assessment may correspond to, or is proportional to, or monotonously increase the IC ₅₀ action of normal cells.

For example, a set of particles can be administered in vivo to neoplastic cells and normal cells in a test animal. For example, the test animal may be a mammal, a primate, a mouse, a rat, a guinea pig, a rabbit or a dog. For example, the efficacy assessment may correspond to the effect of volume reduction of neoplastic cells after administration of the set of particles, or may be proportionally or monotonically increased. For example, the toxicity assessment may correspond to the action of lump reduction of the test animal after administration of the set of particles, or may be proportional thereto or monotonously increased. For example, a set of particles may be administered to a human in clinical trials. The method of treatment of a neoplasm comprises administering a therapeutically effective amount of a compound according to formula I, a compound 1, a polymorph of compound 1 and / or a substantially pure form of the compound 1 to a neoplasm, a mammal or an animal &Lt; / RTI &gt; Prior to administration of the compound, the compound of formula I, the compound 1, the polymorph of compound 1 and / or the substantially pure form of the compound 1 to an animal or human or to a cell in vitro, the particle may be a pharmaceutically acceptable excipient &Lt; / RTI &gt;

An efficiency assessment and / or a toxicity assessment of each set of particles having a first particle size distribution may be made by evaluating the efficiency and / or toxicity of another set or sets of particles having a particle size distribution different from the first particle size distribution And can be compared with the evaluation. A set of particles of a compound having a high efficiency rating and a low toxicity rating may inhibit or kill neoplasms, e.g., cancer, cells, but may be effective to leave normal cells. One of ordinary skill in the art can select an optimal set of particles with a greater efficiency rating, a lower toxicity rating, and / or a larger weighted efficiency rating and a toxicity rating sum, than one or more other sets of particles (e.g., The efficiency evaluation can be weighted with a positive coefficient, and the toxicity assessment can be weighted with a negative coefficient). One of ordinary skill in the art can use another criterion to select particles, for example, an optimal set of particles with a weighted ratio of efficiency evaluations for toxicity assessment and a sum of weighted efficiency assessments. The particle size distribution of the optimal set of particles can take into account the optimal particle size distribution for the compound tested. The optimal particle size distribution may be different for one compound, for example Compound 1 than another compound, for example Compound 1, but not Compound 1. [ The optimal particle size distribution for a given compound can be determined by administration to in vitro cells, small test animals and large test animals. However, the optimal particle size distribution determined by in vitro or in vivo administration of a given compound to an organism may represent a particle size distribution for another compound or a reasonable starting point for administration to another organism.

The optimal set of compounds of formula I, compound 1, polymorph of compound 1 and / or particles of substantially pure form of compound 1 may be included in a composition for reducing or inhibiting the replication or spread of neoplastic cells.

Example

The following examples are provided to further illustrate various aspects of the present invention. The examples also illustrate useful methods for practicing the invention. These embodiments do not limit the claimed invention.

Example  One: Naphthofuran  Preparation of compounds

The procedure for the preparation of the naphthofuran compound (2-acetylnaphtho [2,3-b] furan-4,9-dione) is summarized below:

Step 1: bromination

3-butene-2-one (451.2 g) is added to a 2 L three neck round bottom flask equipped with a mechanical stirrer, thermometer and addition funnel. Bromine (936.0 g) was added to the addition funnel. The contents in the flask were cooled to -5 DEG C, vigorously stirred, and bromine was added dropwise to the flask over 30 minutes while maintaining the temperature at -5 DEG C. [ The mixture is stirred at -5 [deg.] C for an additional 15 minutes and then divided into four equal portions.

Step 2: Tribromide

Each portion of the mixture with tetrahydrofuran (2,133.6 g) is added to a 22 L four-neck round bottom flask equipped with a mechanical stirrer, thermometer and addition funnel. DBU (1,3 diazabicyclo [5.4.0] undec-7-ene, 222.9 g) is added to the addition funnel. DBU is added dropwise over 30 minutes while vigorously stirring while maintaining the temperature at 0 캜 -5 캜. The mixture is stirred at 0 [deg.] C - 5 [deg.] C for an additional 15 minutes.

Step 3: coupling reaction

Then, 2-hydroxy-1,4-naphthofuran (231 g) is added to the flask. Additional DBU (246.0 g) is added to the addition funnel and then added dropwise to the mixture in the flask at a rate such that the temperature of the reaction mixture does not exceed 40 占 폚. After the addition of DBU is complete, the resulting mixture is stirred overnight at room temperature and a sample of the reaction mixture is taken for HPLC analysis.

Step 4: Crystallization

Water (10.8 L) is added to the reaction mixture, the resulting mixture is cooled to 0 to -3 DEG C for at least 30 minutes, and then filtered by a vacuum filter. The filtered solid is successively rinsed with 5% aqueous sodium bicarbonate (3 L), water (3 L), 1% aqueous acetic acid (3 L) and twice ethanol (2 L 1 L)

Store the rinse solids and let them together from other batches. The combined crude product (28.73 kg) is added to a 500 gallon vessel equipped with a mechanical stirrer, thermometer and condenser along with ethyl acetate (811.7 kg). Under a nitrogen atmosphere, the mixture is heated to reflux (72 ° C) for 2 hours and then filtered through a 10 micron cartridge filter containing an activated carbon layer to remove insolubles.

Rinse the vessel, transfer line and filter with fresh hot ethyl acetate (10 kg). The combined filtrates are cooled to 0-5 &lt; 0 &gt; C, held at this temperature for 2 hours, and then filtered through a 20-inch Buchner filter. The filtered solid product is rinsed with 0-5 ° C ethyl acetate (5.7 kg) and dried under vacuum at 40 ° C to constant weight. The remaining filtrate is reduced in volume by up to 63% by evaporation and the crystallization process is repeated again to produce a second crop of product which is dried under the same conditions as the first crop of product.

The lot of naphthofuran compound is obtained by performing the following procedure. The purity of the compound for the lot is 95.44 area% (HPLC).

Example  2: Naphthofuran  Preparation of compounds

Another procedure for the preparation of the naphthofuran compound (2-acetylnaphtho [2,3-b] furan-4,9-dione) is summarized as follows:

Step 1: bromination

Add MVK (2,160 mL, 26.4 mol) to a 12 L RBF (round bottom flask) (using UV filter to block light) and cool to -9.6 C in a dry ice / acetone bath. Bromine (1,300 mL, 25.3 mol) was slowly added over 2 hours 20 minutes while maintaining T = <-2.6 ° C (T _max ). The resulting yellow mixture was stirred for an additional 28 minutes.

Step 2: Dehydrochlorination

The brominated product from above was added to 72 L of RBF with precooled THF (tetrahydrofuran) (20 L, 5 mL / g HNQ (2-hydroxy-1,4-naphthoquinone) -4.8 < 0 &gt; C. DBU (4,200 mL, 28.1 mol) dissolved in THF (4,200 mL) was slowly added over 2 h 20 min while maintaining T <0.3 ° C (T _max ). The resulting suspension was stirred for 42 minutes.

Step 3: Coupling

2-Hydroxy-1,4-naphthofuran (4,003 g, 23.0 mol) was added to the reaction mixture from above at -1.8 [deg.] C in one go. A second portion of the DBU (3,780 mL, 25.3 mol) is added over 48 minutes while the cooling bath is added to bring the reaction temperature to 40 占 폚. The cooling bath was removed, and the reaction mixture was stirred over the weekend over the open air.

Step 4: Isolation of crude

The reaction mixture from above was added to a 200 L reactor with precooled water (100 L, 25 mL / g HNQ). The resulting suspension was cooled to 6.0 占 폚 and stirred at T = 3 占 폚 for about 1 hour. The resulting suspension was then filtered and the collected solid was transferred back to the 200 L reactor.

After stirring in 5% NaHCO ₃ aqueous (26 L, 6.5 mL / g HNQ) for 1 h, the suspension was filtered. The collected solids were transferred to a 200 L reactor and stirred in water (26 L) for 1 hour and then filtered.

The wet solid was again transferred to a 200 L reactor and stirred in 1% aqueous acetic acid (26 L) for about 1 hour, filtered and then washed with water (10 L) on a filter funnel. The collected solids were again transferred to a 200 L reactor and heated to gentle reflux (77.4 C) in ethanol (17.5 L; 4.3 mL / g HNQ). The resulting suspension was cooled to 4.2 &lt; 0 &gt; C and filtered.

The wet solid was transferred to a 100 L reactor and heated to reflux (77.6 C) in ethanol (17.5 L; 4.3 mL / g HNQ). The resulting suspension was cooled to 4.5 &lt; 0 &gt; C and filtered. The wet cake was subjected to de-liquor treatment overnight. ¹ H NMR and HPLC samples were taken. ¹ H NMR: Compound 1 / NDHF (2-acetyl-2,3-dihydronaphtho [2,3-b] furan-4,9-dione) 42:58%; HPLC: Compound 1 / NDHF 74:11 area%.

The solids were dried in a vacuum oven at 50 &lt; 0 &gt; C over 4 days to give 2,268 g of crude compound 1. ¹ H NMR: compound 1 / NDHF 41:59%; HPLC: Compound 1 / NDHF 67:11 area%.

Step 5: Naphthodihydrofuran  Oxidation

The crude compound 1 (2.268 kg) was slurried in toluene (77 L). It was added MnO ₂ (9,536 g), and the mixture was heated to gentle reflux. TLC (1: 1 EA: hexane) showed complete reaction after 1 hour.

The reaction mixture was then filtered through preheated celite pad (1,530 g, bottom layer), activated carbon (2,230 g, middle layer) and celite (932 g, top layer). The yellow-orange filtrate was collected.

The filtrate was concentrated to about 1/10 volume in a rotary evaporator. The slurry was filtered and washed with toluene. The crystals were then dried at 50 &lt; 0 &gt; C to yield 952 g (42%) of a dark yellow solid. HPLC: 99.94%. ¹ H NMR showed no naphthodihydrofuran.

The crystals were dried at 50 &lt; 0 &gt; C under vacuum for an additional 46-65 hours to reduce the amount of residual toluene in the material.

Step 6: Ethyl acetate treatment

Compound 1 (5,816 g) was placed in a 200 L reaction vessel. Ethyl acetate (145 L, 25 mL / g) was added and the solution was heated to reflux over 2 h 26 min. Reflux was maintained for 5 hours 30 minutes, then the mixture was cooled and maintained at 17 [deg.] C overnight.

The slurry was filtered over a polyethylene frit. The yellow crystals were air dried and then placed in a tray in a vacuum oven for 75 hours to give 5,532 g (95.1% yield) of a yellow solid. HPLC: 99.86%. ^& Lt; ¹ &gt; H NMR conforms to the structure of compound 1.

Step 7: Ethyl acetate recrystallization

To 2 L RBF was added the crude (10 g) and ethyl acetate (900 mL). The mixture was refluxed at about 77 DEG C and then ethyl acetate (100 mL) was further added to complete dissolution. The resulting clear yellow solution was heated to reflux for about 30 minutes and then the heating was removed. The mixture was stirred overnight at room temperature.

The resulting suspension was filtered and the collected yellow solid was rinsed with ethyl acetate (30 mL) on a funnel. The wet solid was dried in a vacuum oven at 40-50 &lt; 0 &gt; C over 4 hours to give 8.53 g of yellow crystalline product (total yield about 17%).

¹ H NMR: consistent with structure; HPLC: 99.94 area%; DSC: 228.68 캜, 151 J / g.

Example  3: Naphthofuran  Undifferentiation of compounds

For example, compound 1 crystals were milled and passed through a 160 micron (μm) sieve (sieve # 100, 150 μm aperture) to produce crystals of about 160 microns or less.

For example, Compound 1 crystals were milled to a median particle size of about 20 microns (The Retsch Ultra Centrifugal Mill ZM 200, single pass, at 18,000 rpm using a 0.25 mm screen). Table 3 below shows the distribution of particle sizes (Malvern 2000 with a Hydro 2000S wet accessory). The columns in the table indicate the maximum size of the particles at the cumulative rate indicated by the subscripts in the column headings. For example, column D ₉₀ represents a size where 90% of the particles have the same or lesser size. Column D ₅₀ represents the median size, half of the particles have a larger size, and half of the particles have the same or smaller size.

[Table 3]

For example, Compound 1 crystals were micronized using a jet milling method with a median particle size of about 2 microns as shown in Table 4 below (4 "jet mill, Venturi pressure = 40, milling pressure = 100, feed Speed = 1,304 g / hr.) Particle size analysis was performed using a dry particle method (Sympatec Helos / KF Particle Size Analyzer).

[Table 4]

로서 나타냈으며, 여기서 erf는 오차 함수이며, d는 입자 직경 변수이며, d_중앙은 중앙 입자 크기이며, σ는 누적 분포 함수의 폭(breath)과 관련된 변수이다. CDF(d)는 d 이하의 크기를 갖는 입자의 분율을 나타낸다. d_중앙을 2.07 마이크론의 관찰된 중앙값으로 설정하고, 모델을 대입하여 σ=1.06의 값을 얻었다. 모델은 3.6 마이크론의 평균 직경 및 0.67 마이크론의 모드 직경을 나타냈다. 모델은 또한 표면 거칠기 등의 요인을 고려하지는 않았으나, 2,200 ㎡/㎏의 입자의 비표면적을 시사한다.The cumulative distribution function derived from the log-normal model of the particle size distribution provided an excellent assignment to the data presented in Table 4. The cumulative distribution function

Showed a, where erf is the error function, d is the particle size parameter, d is _{the center} and the center grain size, σ is a parameter associated with the width (breath) of the cumulative distribution function. CDF (d) represents the fraction of particles having a size of d or less. d _center was set at the observed median value of 2.07 microns, and the model was substituted to obtain a value of σ = 1.06. The model showed an average diameter of 3.6 microns and a mode diameter of 0.67 microns. The model also does not take into account factors such as surface roughness, but it suggests the specific surface area of 2,200 m 2 / kg of particles.

Example  4: HPLC  black

This HPLC method is for assessing the purity of naphthofuran, for example 2-acetylnaphtho [2,3-b] furan-4,9-dione (Compound 1) and its reaction completion by HPLC. All components are expressed as percent area of all peaks in the chromatogram.

1. Devices and Materials

[Table 5A]

2. Solution Manufacturing

10 mM  phosphate Buffer

Weigh 1.74 g of potassium phosphate, dibasic and dilute with 1 L of purified water (adjust weight and volume for the required amount). The pH is adjusted to pH 6.8 using phosphoric acid.

Mobile phase  A

10 mM phosphate buffer and acetonitrile are mixed at 80:20 buffer: acetonitrile ratio to produce mobile phase A. Degassing.

Mobile phase  B

10 mM phosphate buffer and acetonitrile are mixed in a 20:80 buffer: acetonitrile ratio to produce mobile phase B. Degassing.

diluent

Mobile phase A will be used as a diluent for all samples and standard formulations.

3. Standard formulation

1.0 ㎎/㎖) Compound 1 Stock standard (concentration

1.0 mg / ml)

10 mg of Compound 1 reference material was weighed into a 20 ml scintillation vial; Weight &lt; RTI ID = 0.0 &gt; +/- 0.01 &lt; / RTI &gt; 10 ml of DMSO is added and sonicated until the solids are dissolved.

1.0 ㎎/㎖) Stock test sample (concentration

1.0 mg / ml)

A 10 mg sample is weighed into a 20 ml scintillation vial and diluted with 10 ml DMSO to generate a test solution.

Working test sample (concentration 0.01 mg / ml)

Transfer 1 ml to a 100 ml volumetric flask and dilute with a diluent solution to form a solution.

4. Operating conditions

[Table 5B]

5. Operation procedure

The solution is injected in the following order:

1. Thinner Blank (1X)

2. Compound 1 working standard (5X)

3. Test solution (2X each)

4. Working standards (1X each)

6. System Suitability

The system is suitable for use when it meets the following criteria.

1. The diluent blank injection at the beginning of the procedure does not contain peaks that are disturbed by any identified impurities.

2. The initial 5 repeated injections of the compound 1 working standard resulted in: (1)% RSD _{peak area} < 3.0%; (2)% RSD _{retention time} < 3.0%; And (3) an average tailing factor < 2.0.

3. In the chromatogram for the parenthesized standard, (1) the retention time is 97.0-103.0% of the mean retention time from the initial conformity injection, and (2) its area% is 97.0-103.0% of the initial value.

7. Calculation

All peaks will be reported as area% of the total peak in the chromatogram, which will be calculated by the integration software according to the following equation.

NMR and TLC

[Table 5C]

[Table 5D]

Example 5 : Preparation of crude 2-acetylnaphtho [2,3-b] furan-4,9-dione

Another procedure for the preparation of Compound 1 is summarized as follows.

Bromine (0.95 eq.) Is added to the methyl vinyl ketone (MVK, 1.0 eq.) At -20 to -15 <0> C via addition funnel while maintaining the reaction temperature below 0 <0> C. After stirring the reaction mixture at -10 to 0 C for an additional 2 to 3 hours, tetrahydrofuran (6 vol) is added and the reaction mixture is cooled to -20 to -10 [deg.] C. Triethylamine (1.1 eq.) Is then added with vigorous stirring while maintaining the reaction temperature below 0 &lt; 0 &gt; C. The resulting slurry is stirred at -15 to -5 [deg.] C for a minimum of 10 hours, then warmed to -5 to 5 [deg.] C and filtered. The filtrate is then analyzed by in-process ¹ H NMR to determine the amount (wt%) of the intermediate bromomethyl vinyl ketone (BrMVK) present and maintained at -25 to -10 ° C until further use.

Then, 2-hydroxy-1,4-naphthoquinone (1.0 equivalent to the calculated amount of BrMVK from in-process ¹ H NMR) is added to tetrahydrofuran (3.15 vol) in a clean reaction vessel. The resulting orange slurry is briefly stirred and then 1,8-diazabicyclo [5.4.0] undec-7-ene (DBU, 1.1 eq.) Is added while maintaining the temperature below 45 ° C. The reaction mixture is then stirred at 40 to 45 캜 for a minimum of 1 hour, heated to 50 to 55 캜, and the BrMVK solution is added by addition funnel while maintaining the reaction temperature at 50 to 60 캜. The reaction mixture is then stirred at 50 to 55 캜 for about 18 hours until less than 5% of 2-hydroxy-1,4-naphthoquinone remains. The reaction mixture is then concentrated, co-evaporated twice with ethanol and recrystallized from ethanol / water (1: 1). The solids are dried under vacuum at 35-45 [deg.] C. The crude solid and Charcoal G-60 (100 wt%) are suspended in acetonitrile, heated at 70-75 [deg.] C for 2 hours, filtered and washed with hot acetonitrile. The filtrate is then concentrated to 1/3 volume, cooled to 0-5 &lt; 0 &gt; C and filtered. The solid is then dried under vacuum at 45-50 &lt; 0 &gt; C. The crude solid is then slurried again in ethyl acetate at reflux for 6 hours, cooled to room temperature, filtered and washed with ethyl acetate. The material is then dried under vacuum at 45-50 ° C and packaged for final release.

Example  6: Clinical trials: safety and efficacy

The dose of the compound 2-acetylnaphtho [2,3-b] furan in order to enter the Phase I clinical trial after receiving the IND approval from the US FDA and the Canadian Department of Health, 4,9-dione. Each cycle consists of oral administration of compound twice a day for 4 weeks. The cycle was repeated every 4 weeks (28 days) until progression of the disease, unacceptable toxicity or other discontinuity criteria was met. The dose escalation test was performed as an open label and multi-centered test. A modified Simon accelerated titration reaction was used for dose escalation.

The primary goal of the study was to determine safety, tolerability, and recommended Phase II dosing (RP2D). The secondary objective of the test is to measure the pharmacokinetic profile of the compound, the pharmacodynamics of the compound and the pre-antitumor activity of the compound.

Included criteria are metastatic, non-resectable or relapsing histologically or cytologically confirmed solid tumors; ≥ 18 years of age; Measurable disease by RECIST; And Karnofsky ≥70%. Exclusion criteria include chemotherapy, radiotherapy, immunotherapy or research drugs within 4 weeks of the first administration; Surgery within 4 weeks of initial administration; And a known brain metastasis.

Demographic and baseline disease characteristics of selected patients under the above criteria are summarized in Table 6 below.

[Table 6]

Among these patients, 10 cohorts were evaluated at doses ranging from 20 mg to 2,000 mg / day. No dose limiting toxicity was observed. The most common adverse events were diarrhea, nausea and fatigue. Examples of Grade 3 or higher include fatigue and diarrhea. Adverse events are summarized in Table 7 below.

[Table 7]

At 20 mg daily dosing, a surprisingly high concentration of compound was observed in the patient &apos; s urine. Moreover, Applicants have tested the antitumor activity of the compounds of the invention in patient urine, and the compounds have been found to remain efficacious against cancer cells.

Among the treated patients, the disease control (disease stabilization and tumor regression) is a tumor response in various tumors that are refractory to chemotherapy including colorectal adenocarcinoma, head and neck cancer, breast cancer, stomach cancer, ovarian cancer, chondrosarcoma, adrenocortical carcinoma and melanoma In 65% of patients who can be evaluated. There was one complete regression of kidneys of colon cancer metastatic lesions (patient 0001). Patients treated with Compound 1 showed a dramatic lack of new metastatic tumor lesions. Showed no more than 80% metastatic tumors among 24 evaluable patients with advanced refractory cancers.

Patients enrolled for signs of activity are summarized in Table 8 below.

[Table 8]

Applicants have also found that a high level of p-STAT3 in tumor tissue predicts an excellent response of the tumor to Compound 1 prior to treatment with immunohistochemistry using anti-p-STAT3 antibody.

The pharmacokinetic profile of oral BID administration was also tested. Plasma concentrations of the drug were achieved several times over an effective concentration (IC _{50 in} vitro) as illustrated in Table 9 below. However, the drug concentration was not maintained at a high level for a long time, and was rapidly reduced below the effective concentration.

[Table 9]

To maintain the plasma plasma concentration at or above the effective level for a desired period of time and to further increase the peak plasma concentration, Applicants have developed 500 (between 4 doses of the same day or "q4h & The pharmacokinetics of ㎎ BID therapy were studied and compared to those of 500 ㎎ QD regimen (Fig. 1). Surprisingly, no significant difference was observed with respect to pharmacokinetics between the two dosing regimens. Previously, using BID therapy, it was expected that drug levels in the patient plasma would show another distinct peak as dosing would be doubled in the same day as compared to QD therapy. However, the twice q4h administration of Compound 1 failed to maintain drug levels for the same length of time after the first administration for the same 24 hour period. In another suitable dose regimen, 500 mg of Compound 1 was administered to a human subject three times daily (TID). Somewhat disappointingly, the level of patient exposure to Compound 1 was not significantly improved by three doses per day compared to twice daily dosing.

Example  7: Administration regimen and new formulation

A therapeutically effective amount of a pharmaceutical composition comprising a particle, polymorph and / or purified form of a compound of the present invention may be in the range of about 160 mg to about 1,000 mg, for example a total daily dose of about 960 mg . However, in order to achieve an effective dosage level, clinical trials have faced the challenge of patient burden. Higher strength capsules were designed into the new formulation (DP2A) in order to overcome the pill burden and to solve the problem of maintaining drug concentration at the minimum effective level or higher for the desired period of time.

However, with the use of higher strength capsules, the Applicant observed adverse gastrointestinal effects, including nausea, vomiting and diarrhea in patients. When Applicants study the relationship between plasma drug concentrations and adverse effects in additional Phase I clinical trials, some of them are summarized in Table 10 below and, surprisingly, the data indicate that the severity of gastrointestinal adverse events is as suspected (See Tables 10 and 11 below). &Lt; tb &gt; &lt; TABLE &gt;

[Table 10]

[Table 11]

After a difficult conventional solution, breakthroughs result from unexpected changes in the drug intake protocol. Surprisingly, Applicants have found that the interval between doses according to some preferred embodiments of the present invention is a key factor for reduced gastrointestinal side effects as well as prolonged drug exposure. Even more surprisingly, instead of concentrating the drug administration by shortening the interval between each ingestion as intuitively attempted when a drastic reduction in drug concentration in the bloodstream is questioned, Applicants have found that extending that interval is indeed a problem . For example, the preferred interval between administration of the drug is from about 8 hours to about 14 hours, more preferably from about 10 hours to about 13 hours. In certain embodiments, the compounds or related compositions and forms of the invention are administered twice daily at an average interval of about 12 hours over the period between administrations wherein each dose is about 480-500 mg BID.

In another suitable administration regimen, more than about 20 mg or 20 mg of Compound 1 is administered once a day to a human subject. These dosing regimens, referred to herein as 20 mg QD, have been shown to be therapeutically active levels in patients, but the drug is rapidly cleared from human blood. However, as the drug was cleared from the bloodstream through the kidneys into the urine, very high concentrations of drug in the urine showed signs of antitumor activity particularly effective in kidneys with colon cancer lesions. In general, such dosing regimens have demonstrated excellent human tolerability.

In another suitable administration regimen, Compound 1 is administered with a fluid, such as milk or water, on an empty bed improving the pharmacokinetic exposure (Table 12 below). Semi-intuitively, milk helps patients with gastrointestinal adverse effects.

[Table 12]

In another suitable dosing regimen, Compound 1 was administered with food to delay Tmax (Table 13 below).

[Table 13]

In another suitable dose regimen, the pill burden problem was addressed by the new drug formulation (DP2A). The new formulation replaces most of the surfactant Gel Lucir ™ 44/14 used in the DP1 formulation with another surfactant Labrafil and reduces the capsule size from size 00 to size 1 or size 2, do. The new formulation was able to maintain similar bioavailability (Figure 2). The components of the two formulations are summarized below (Table 14 below):

[Table 14]

Additional studies were conducted using oral formulations of the compounds of the present invention, specifically higher strength capsules (DP2A). As described herein, the compounds of the present invention inhibit cancer stem cell (CSC) self-renewal, inhibit the Stat3, beta -catenin and Nanol pathway and induce apoptosis in CSC as well as non-stem cancer cells , And showed effective antitumor and anti-metastatic activity before clinical use. In the Phase I studies described above, the compounds showed tolerability as well as an indication of anticancer activity in patients with solid tumors. The study described herein was designed as an amplification step 1 for evaluating drugs designed for pivotal testing to measure pharmacokinetics (PK) in patients with advanced cancer.

On day 1, the patient administered a single 500 mg dose of the oral dosage form of the compounds of the invention (DP1). On days 4 and 8, a higher strength capsule (DP2A) for pivotal testing was given with fasting and then the condition was provided. Thereafter, DP2A was administered daily until disease progression or unacceptable toxicity occurred. Endpoints were safety, PK and pre-anticancer activity.

DP2A was evaluated in 24 patients. There was no significant difference in plasma exposure between DP1 and DP2A and no significant food effect was observed. Nine patients received 500 mg of compound DP2A twice daily (DP2A-4h) twice daily and 15 patients received compound DP2A 500 mg BID every 12 hours (DP2A-12h). Despite the equivalence to compound DP1, DP2A-4h has a higher frequency of gastrointestinal (GI) adverse events (AE) than those observed in the prior art experiments described above, including diarrhea, convulsive abdominal pain, zone / vomiting, . Conversely, DP2A-12h has less GI AE and was selected for dilation experiments. Out of 15 patients who received DP2A-12h, 8 CRC patients were enrolled and disease control was 67%, which was assessable for response (4/6) with progression-free survival and overall survival at 17 and 39 weeks, respectively .

The recommended dosing regimen for the compounds in the pivotal study was determined to be about 500 mg BID q12h. Signs of anticancer activity were observed in patients with CRC and ovarian cancer.

Example  8: Anti-mitotic agent  Used combination therapy

The compounds of the invention have been used in combination with antimitotic agents that have proven to be particularly effective chemotherapeutic agents for the successful treatment of patients. Examples of antimitotic agents that may be useful in combination therapy with the compounds of the present invention include paclitaxel (ablucan / taxol), docetaxel (taxotere), BMS-275183, xyotax, tocosal, vinorelbine, , Vinblastine, vindesine, binzolidine, etoposide (VP-16), tenniposide (VM-26), ixabepilone, laurotaxel, But is not limited thereto.

The Phase Ib experiment was designed to evaluate the combination of the compound of the present invention and paclitaxel in patients with advanced malignant tumors. The experiment was designed as a step Ib dose-increase experiment to measure the safety, tolerability, RP2D and pre-anticancer activity of the compounds of the invention when used with paclitaxel per week. The compound was administered in three increasing dose cohorts (200 mg BID, 400 mg BID, 500 mg BID) until the progress of the disease, unacceptable toxicity or other discontinuity criteria was met, 3 weeks for 4 weeks).

Twenty-four patients were enrolled in this study. The compound monotherapy RP2D of the present invention can be administered in combination with paclitaxel at a total dose. The maximum internal dose (MTD) was not determined. No new adverse events were observed, and the safety profile was similar to that of each drug as a monotherapy. The most common adverse events included grade 1 and 2 diarrhea, convulsive abdominal pain, nausea, and vomiting. The grade 3 case involved protocol therapy in 4 patients, including diarrhea, dehydration and weakness. No significant pharmacokinetic interactions were observed. Disease control (ie, the sum of complete response (CR) + partial response (PR) + stable disease (SD)) was observed in 10 of 15 (67%) evaluable patients. Five patients with refractory gastrointestinal adenocarcinoma (GEJ) adenocarcinoma were enrolled, two were PR (48% and 44% degenerative), one was SD with 25% degeneration, Two (previously failing taxa) were extended to SD ≥24 weeks.

These Phase Ib experiments have demonstrated that the compounds of the present invention and paclitaxel per week can be safely combined at full dose. An encouraging antitumor activity was observed in patients with stomach and GEJ adenocarcinoma.

[Table 15]

Phase II trials are ongoing in magnification from the Phase Ib trial and continue to register patients with gastric / GEJ adenocarcinoma.

[Table 16]

In these experiments, seven out of nine evaluable gastric / GEJ patients exhibited activity in response to the combination of the compound of the present invention and paclitaxel.

Example  9: Predictability As a biomarker  P- STAT3

Archival tumor tissue samples of CRC patients were analyzed by immunohistochemistry (IHC) using antibodies labeled against phosphorylated STAT3 (p-STAT3). As shown in FIG. 3B, the compounds of the present invention were highly effective in suppressing p-STAT3 expression. There was no longer any detectable p-STAT3 in patient tissues after treatment, even at doses as low as 100 QD (single daily dose). As the chart of FIG. 3b shows, for patients receiving the compound treatment of the invention (N = 13), overall survival (OS) is much more optimistic for patients who have already displayed relatively high levels of p-STAT3. For example, only 40% of patients with high p-STAT3 levels after treatment survived longer than 10 weeks, whereas only 10% of patients with low p-STAT3 levels before treatment or no p-STAT3 levels Survived more than 100 weeks. This further confirms that the compounds of the present invention down-regulate the STAT3 pathway and that the STAT3 pathway is associated with colorectal cancer.

A direct correlation between p-STAT3 levels and OS in CRC patients treated with the compounds of the invention produces p-STAT3, a promising diagnostic biomarker that can be used to predict therapeutic efficacy. Thus, p-STAT3 levels may be used to screen patient pools for treatment with the compounds of the present invention.

Example  10: Predictability As a biomarker  Nuclear β- Catechin

Archival tumor tissue samples of CRC patients were analyzed by immunohistochemistry (IHC) using labeled antibodies to [beta] -catenin. As shown in Figure 4a, the compounds of the present invention are effective in eliminating or preventing the accumulation of beta -catenin in the nucleus of a tumor tissue. As shown in the chart in FIG. 4B, in the case of patients receiving treatment with a compound of the present invention (N = 13), overall survival (OS) has already been found to indicate a high level of pre-treatment nuclear beta -catenin , Much more optimistic. For example, nearly 40% of patients with high nuclear beta -catenin levels prior to treatment survived longer than 100 weeks, while patients with high levels of membrane beta -catenin did not survive beyond 25 weeks. This further confirms that the compounds of the present invention disrupt or modulate the? -Catenin action and that the? -Catenin pathway is associated with colorectal cancer.

A direct correlation between nuclear beta -catenin levels and OS in treated CRC patients using compounds of the invention produces nuclear beta -catenin levels, which are promising diagnostic biomarkers that can be used to predict therapeutic efficacy. Thus, nuclear beta -catenin levels may be used to screen patient pools for treatment with the compounds of the invention.

Example  11: Compound of the present invention In vitro  And In vivo  analysis

CD44 ^solid cells were isolated by FACS (FaDu) and their growth was blocked by the compounds of the invention (Figure 5).

And, in vivo experiments of nude mice with xenografted colon cancer tumor tissues showed that the compounds of the present invention were effective in reducing or cleaning p-STAT3 and beta -catenin levels (Fig. 6).

Mouse experiments also showed that the compounds of the present invention target cancer stem cells (Figure 7).

In human clinical trials, the compounds of the present invention were found to be effective in CRC patients (Figure 8).

The embodiments illustrated and discussed herein merely teach those skilled in the art the best method known to the inventors for creating and using the present invention. Nothing herein is to be construed as limiting the scope of the present invention. All of the embodiments presented are for purposes of illustration and not limitation. The above-described embodiments of the present invention may be modified or changed without departing from the invention as will be appreciated by those skilled in the art in light of the above teachings. It is therefore to be understood that within the scope of the claims and their equivalents the invention may be practiced other than as specifically described.

Claims (119)

A method of treating cancer in a human subject comprising administering to a subject in need thereof a therapeutically effective amount of the following structure:

Or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, wherein said compound is administered to said subject in a total daily dosage within the range of from about 80 mg to about 2,000 mg, How it is. 10. The method of claim 1, wherein the compound is administered to the subject at a total daily dose of about 960 mg. 3. The method according to claim 1 or 2, wherein the compound is administered to the subject in a twice-a-day dose. 4. The method of claim 3, wherein each dose is about 480 mg. 2. The method of claim 1 wherein said compound is administered to said subject as a tablet or capsule. 6. The method of claim 5, wherein the tablet or capsule comprises a dose of about 80 mg. 7. The method of any one of claims 1-6, wherein the interval between administration of the compound is in the range of about 4 hours to about 16 hours. 8. The method of claim 7 wherein said compound is administered in an amount selected from the group consisting of about 80 mg BID, about 160 mg BID, about 320 mg BID, about 400 mg BID, about 480 mg BID, about 500 mg BID, &Lt; / RTI &gt; to said subject. 7. The method of any one of claims 1 to 6 wherein the interval between administrations of the compound is at least about 4 hours, at least about 5 hours, at least about 6 hours, at least about 7 hours, at least about 8 hours, at least about 9 hours About 10 hours or more, about 11 hours or more, about 12 hours or more, about 13 hours or more, about 14 hours or more, about 15 hours or more, and about 16 hours or more. 7. The method according to any one of claims 1 to 6, wherein the interval between doses is about 12 hours. 2. The method of claim 1 wherein each dose is about 480 mg BID and the interval between doses is about 12 hours. 2. The method of claim 1 wherein each dose is about 80 mg BID and the interval between doses is about 12 hours. 2. The method of claim 1 wherein each dose is about 400 mg BID and the interval between doses is about 12 hours. 2. The method of claim 1 wherein each dose is about 320 mg BID and the interval between doses is about 12 hours. 2. The method of claim 1, wherein said compound is orally administered in combination with a fluid to an empty stomach. 16. The method of claim 15, wherein the fluid comprises milk or water. The compound according to claim 1, wherein said compound is
(a) a polymorphism characterized by an X-ray diffraction pattern substantially similar to that shown in Figure 2 of patent publications WO 2011/116398 and WO 2011/116399;
(b) at least about 10.2, 11.9, 14.1, 14.5, 17.3, 22.2, and 28.1 degrees two theta peaks, -Naphtho [2,3-b] furan-4,9-dione;
(c) a peak at at least about 10.2 degrees two theta, at least about 11.9 degrees two theta peak, at least about 14.1 degrees two theta peak, at least about 14.5 degrees two theta peak, at least about 17.3 degrees two theta peak, Acetyl-4H, 9H-naphtho [2, &lt; / RTI &gt; 2] pyridine characterized by an X- ray diffraction pattern comprising at least two peaks from a peak at 22.2 degrees two theta and a peak at least about 28.1 degrees two theta, , 3-b] furan-4,9-dione;
(d) a polymorph characterized by an X-ray diffraction pattern substantially similar to that shown in Figure 3 of patent publications WO 2011/116398 and WO 2011/116399;
(e) a peak at at least about 7.5, 9.9, 12.3, 15, 23, 23.3, 24.6 and 28.4 degrees two theta, 4H, 9H-naphtho [2,3-b] furan-4,9-dione; And
(f) a peak at least about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 12.3 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 23 degrees two theta, Characterized by an X-ray diffraction pattern comprising two or more peaks from a peak at 23.3 DEG 2 &amp;thetas; a peak at least at about 24.6 DEG 2 &amp;thetas; and a peak at least about 28.4 DEG 2 theta, -4H, 9H-naphtho [2,3-b] furan-4,9-dione
&Lt; / RTI &gt; 18. The method according to any one of claims 1 to 17, wherein the compound is present in the form of particles and the particles have a diameter of about 20 占 퐉 or less. The method of claim 1, wherein said compound is present in a pharmaceutical composition comprising a population of said compound particles, and wherein a portion of the cumulative total of said particles has a diameter in the range of 0.2 탆 to 20 탆. The method of claim 1 wherein said compound is present in a pharmaceutical composition comprising a population of said compound particles and wherein 50% (D50) of the cumulative total of said particles has a diameter in the range of from about 0.5 to about 5 micrometers . 2. The method of claim 1, wherein said compound is present in a pharmaceutical composition comprising a population of said compound particles and wherein 50% (D50) of the cumulative total of said particles has a diameter of about 2 microns. 22. The compound according to any one of claims 1 to 21, wherein the compound is an API having a purity of at least 95.0% as determined by high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR) Wherein the composition comprises no more than 5% of impurities. The method according to claim 1, wherein the cancer is selected from the group consisting of gastric and gastric adenocarcinoma, colorectal adenocarcinoma, breast cancer, ovarian cancer, head and neck cancer, melanoma.
21. The method of claim 1, wherein the cancer is gastric cancer. 8. The method of claim 1, wherein the cancer is a gastric / gastrointestinal adenocarcinoma. The method of claim 1, wherein the cancer is refractory. The method of claim 1, wherein the cancer recurs. 2. The method of claim 1, wherein the cancer is metastatic. 2. The method of claim 1, wherein said cancer is associated with overexpression of STAT3. 2. The method of claim 1, wherein the method further comprises detecting a level of phosphorylated STAT3 (p-STAT3) in the patient tissue, wherein the level of p-STAT3 is used as a biomarker for patient selection. 2. The method of claim 1, wherein said cancer is associated with nuclear beta -catenin overexpression. 2. The method of claim 1, wherein said method further comprises detecting nuclear beta -catenin expression in a tissue of a patient, wherein said nuclear beta -catenin expression is used as a biomarker for patient selection. As a method for treating a curable or prophylactic cancer,
(a) for a subject in need of a therapeutic or prophylactic cancer treatment, the following structure:

Or a pharmaceutically acceptable salt, solvate, hydrate, or prodrug thereof; And
(b) administering to said subject a dose of an anti-mitotic agent, or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof,
&Lt; / RTI &gt; CLAIMS 1. A method of treating a cancer, either healing or prophylactically, by administering to a subject paclitaxel (ablacic acid / taxol)
(a) for a subject in need of a therapeutic or prophylactic cancer treatment, the following structure:

Or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, wherein the dose of the first agent is selected from the group consisting of paclitaxel (ablucan / taxol) Is administered before or after administration to the subject. 33. The method of claim 32, wherein said anti-mitotic agent is selected from the group consisting of paclitaxel (ablucan / taxol), docetaxel (taxotere), BMS-275183, xyotax, tocosal, vinorelbine, vincristine, vinblastine, Wherein the cell is selected from the group consisting of zolyidins, ethoposide (VP-16), tennifoside (VM-26), xabaperolone, laurotaxel, 34. The method of claim 33, wherein the anti-mitotic agent comprises paclitaxel (ablucan / taxol). 34. The method according to claim 32 or 33, wherein the cancer is selected from the group consisting of gastric cancer, gastrointestinal cancer and esophageal cancer. 34. The method of claim 32 or 33 wherein said cancer is gastric / gastric adenocarcinoma. 34. The method of claim 32 or 33, wherein the cancer is refractory. 34. The method of claim 32 or 33, wherein the cancer recurs. 34. The method of claim 32 or 33, wherein the cancer is metastatic. 33. The method of claim 32 or 33, further comprising detecting a level of phosphorylated STAT3 (p-STAT3) in the patient's tissue, wherein the level of p-STAT3 is used as a biomarker for patient selection. 33. The method according to claim 32 or 33, wherein said cancer is associated with overexpression of nuclear beta -catenin. 33. The method of claim 32 or 33, further comprising detecting nuclear beta -catenin expression in the tissue of the subject, wherein nuclear beta -catenin expression is used as a biomarker for patient selection. A method of treating cancer in a selected patient population,
(a) measuring the level of phosphorylated STAT3 (p-STAT3) in a biological sample obtained from a patient candidate diagnosed with cancer;
(b) confirming that the p-STAT3 level of the patient candidate is higher than a benchmark level; And
(c) a therapeutically effective amount of the following structure:

Or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof.
&Lt; / RTI &gt; 45. The method of claim 44, wherein said cancer is selected from the group consisting of esophageal cancer, gastric adenocarcinoma, glioma adenocarcinoma, chondrosarcoma, colon cancer, colon adenocarcinoma, rectal adenocarcinoma, colon adenocarcinoma, breast cancer, ovarian cancer, head and neck cancer, melanoma, Lt; RTI ID = 0.0 &gt; neoplastic &lt; / RTI &gt; carcinoma. 45. The method of claim 44, wherein the cancer is a colon adenocarcinoma. 45. The method of claim 44, wherein the cancer is refractory. 45. The method of claim 44, wherein the cancer recurs. 45. The method of claim 44, wherein the cancer is metastatic. A method of treating cancer in a selected patient population,
(d) detecting nuclear β-catenin expression in a biological sample obtained from a patient candidate diagnosed with cancer;
(e) confirming that significant beta -catenin expression is detected in the nuclei in the sample from the patient candidate; And
(f) a therapeutically effective amount of the following structure:

Or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof.
&Lt; / RTI &gt; 53. The method of claim 50, wherein the cancer is selected from the group consisting of gastric and gastric adenocarcinoma, esophoid adenocarcinoma, colorectal adenocarcinoma, breast cancer, ovarian cancer, head and neck cancer, melanoma and adrenocortical carcinoma. 51. The method of claim 50, wherein the cancer is a colon adenocarcinoma. 51. The method of claim 50, wherein the cancer is refractory. 51. The method of claim 50, wherein the cancer recurs. 51. The method of claim 50, wherein the cancer is metastatic. A method of treating cancer in a human subject comprising administering to a subject diagnosed with cancer a therapeutically effective amount of the following structure:

Or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, wherein the cancer is a colon cancer, a colon adenocarcinoma or a rectal adenocarcinoma. 57. The method of claim 56, wherein said compound is administered to said subject in a dose of twice daily within a total of about 80 mg to about 2000 mg. 57. The method of claim 56, wherein said compound is administered to said subject at a dose of about 480 mg. 57. The method of claim 56, wherein the interval between administration of said compound is in the range of about 4 hours to about 16 hours. 56. The method of claim 56, wherein said compound is administered to said subject at a dose selected from the group consisting of about 80 mg BID, about 160 mg BID, about 320 mg BID, about 400 mg BID, about 480 mg BID, &Lt; / RTI &gt; 57. The method of claim 56, wherein the cancer is refractory. 57. The method of claim 56, wherein the cancer recurs. 57. The method of claim 56, wherein the cancer is metastatic. A therapeutically effective amount of the following structure:

&Lt; / RTI &gt;
Surfactants including sodium lauryl sulfate (SLS) or sodium dodecyl sulfate (SDS);
Gel lucer (lauroyl polyoxyl glyceride); And
Labrafil (linoleoyl polyoxyl glyceride)
&Lt; / RTI &gt; 65. The pharmaceutical composition of claim 64 further comprising about 27.18% by weight of active ingredient, about 0.27% by weight of surfactant, about 14.51% by weight of gel lucer and about 58.04% by weight of labrafil. 65. The pharmaceutical composition of claim 64 further comprising about 125 mg of active ingredient, about 1.2 mg of a surfactant, about 66.8 mg of gel lucer and about 267 mg of labrafil. 65. The pharmaceutical composition of claim 64 further comprising about 80 mg of active ingredient, about 0.8 mg of a surfactant, about 42.7 mg of gel lucer and about 170.9 mg of labrafil. 67. An article of manufacture comprising a pharmaceutical composition of any one of claims 64 to 67 contained within a capsule. 69. The article of manufacture of claim 68, wherein the capsule is size 1 or smaller. A method of treating cancer in a selected patient population,
(a) a level of one or more cancer stemness markers selected from phosphorylated STAT3 (p-STAT3), beta -catenin and NANOG in a biological sample obtained from a candidate patient with cancer and / Measuring subcellular localization;
(b) confirming that the cancer stem cell marker level and / or intracellular localization of the patient candidate is higher than a reference level; And
(c) a therapeutically effective amount of the following structure:

Or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof.
&Lt; / RTI &gt; 70. The method of claim 70, wherein said cancer is selected from the group consisting of colorectal adenocarcinoma, breast cancer, ovarian cancer, head and neck cancer, melanoma, gastric / gastric adenocarcinoma, esophageal adenocarcinoma and adrenocortical carcinoma. 71. The method of claim 70, wherein the cancer is a colon adenocarcinoma. 71. The method of claim 70, wherein the cancer is refractory. 71. The method of claim 70, wherein the cancer recurs. 71. The method of claim 70, wherein the cancer is metastatic. A method of treating cancer in a human subject comprising administering to a subject in need thereof a therapeutically effective amount of the following structure:

Or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof, and administering a therapeutically effective amount of paclitaxel. 77. The method of claim 76, wherein said compound is administered to said subject in a total daily dosage within the range of about 80 mg to about 2000 mg. 77. The method of claim 76, wherein said compound is administered to said subject at a total daily dose of about 960 mg. 78. The method of any one of claims 76-78, wherein the compound is administered to the subject in a twice daily dosage. 80. The method of claim 79, wherein each dose is about 480 mg. 80. The method according to any one of claims 76 to 80, wherein the interval between administration of the compound is in the range of about 4 hours to about 16 hours. 83. The method of claim 81, wherein said compound is administered to said subject at a dose selected from the group consisting of about 80 mg BID, about 160 mg BID, about 320 mg BID, about 400 mg BID, about 480 mg BID, &Lt; / RTI &gt; 80. The method of any one of claims 76 to 80 wherein the interval between administrations of the compound is at least 4 hours, at least 5 hours, at least 6 hours, at least 7 hours, at least 8 hours, at least 9 hours, at least 10 hours, 11 hours or more, 12 hours or more, 13 hours or more, 14 hours or more, 15 hours or more, and 16 hours or more. 80. The method according to any one of claims 76 to 80, wherein the interval between doses is about 12 hours. 77. The method of claim 76, wherein each dose is about 480 mg BID and the interval between doses is about 12 hours. 77. The method of claim 76, wherein each dose is about 80 mg BID, and the interval between doses is about 12 hours. 80. The method of claim 76, wherein each dose is about 400 mg BID, and the interval between doses is about 12 hours. 80. The method of claim 76, wherein each dose is about 320 mg BID and the interval between doses is about 12 hours. 80. The method of claim 76, wherein the compound is orally administered in combination with a fluid to an empty stomach. 77. The method of claim 76, wherein the fluid comprises milk or water. 77. The method of claim 76, wherein the paclitaxel is administered to the subject in a total weekly dosage within the range of about 40 mg / m 2 to about 100 mg / m 2. 77. The method of claim 76, wherein the paclitaxel is administered to the subject in a total weekly dose of about 80 mg / m &lt; 2 &gt;. 77. The method of claim 76, wherein the paclitaxel is administered to the subject via IV. 78. The compound of claim 76, wherein said compound is
(a) a polymorphism characterized by an X-ray diffraction pattern substantially similar to that shown in Figure 2 of patent publications WO 2011/116398 and WO 2011/116399;
(b) at least about 10.2, 11.9, 14.1, 14.5, 17.3, 22.2, and 28.1 degrees two theta peaks, -Naphtho [2,3-b] furan-4,9-dione;
(c) a peak at at least about 10.2 degrees two theta, at least about 11.9 degrees two theta peak, at least about 14.1 degrees two theta peak, at least about 14.5 degrees two theta peak, at least about 17.3 degrees two theta peak, Acetyl-4H, 9H-naphtho [2, &lt; / RTI &gt; 2] pyridine characterized by an X- ray diffraction pattern comprising at least two peaks from a peak at 22.2 degrees two theta and a peak at least about 28.1 degrees two theta, , 3-b] furan-4,9-dione;
(d) a polymorph characterized by an X-ray diffraction pattern substantially similar to that shown in Figure 3 of patent publications WO 2011/116398 and WO 2011/116399;
(e) a peak at at least about 7.5, 9.9, 12.3, 15, 23, 23.3, 24.6 and 28.4 degrees two theta, 4H, 9H-naphtho [2,3-b] furan-4,9-dione; And
(f) a peak at least about 7.5 degrees two theta, a peak at least about 9.9 degrees two theta, a peak at least about 12.3 degrees two theta, a peak at least about 15 degrees two theta, a peak at least about 23 degrees two theta, Characterized by an X-ray diffraction pattern comprising two or more peaks from a peak at 23.3 DEG 2 &amp;thetas; a peak at least at about 24.6 DEG 2 &amp;thetas; and a peak at least about 28.4 DEG 2 theta, -4H, 9H-naphtho [2,3-b] furan-4,9-dione
&Lt; / RTI &gt; 94. The method according to any one of claims 1 to 94, wherein the compound is present in the form of particles and the particles have a diameter of about 20 占 퐉 or less. 77. The method of claim 76, wherein said compound is present in a pharmaceutical composition comprising a population of said compound particles, and wherein a portion of the cumulative sum of particles has a diameter within the range of from about 0.5 占 퐉 to about 5 占 퐉. 77. The method of claim 76, wherein said compound is present in a pharmaceutical composition comprising a population of said compound particles and wherein 50% (D50) of the cumulative total of the particles has a diameter of about 2 占 퐉. 98. The compound according to any one of claims 1 to 97, wherein the compound has an API having a purity of at least 95.0% as determined by high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR) or both HPLC and NMR Wherein the composition comprises no more than 5% of impurities. 98. The method according to any one of claims 1 to 98, wherein the cancer is selected from the group consisting of gastric and gastric adenocarcinoma, breast cancer, ovarian cancer, head and neck cancer, melanoma and esophagus adenocarcinoma. 77. The method of claim 76, wherein the cancer is gastric and gastric adenocarcinoma. 77. The method of claim 76, wherein the cancer is selected from the group consisting of colorectal adenocarcinoma, breast cancer, ovarian cancer, head and neck cancer, melanoma, gastric adenocarcinoma and adrenocortical carcinoma. 77. The method of claim 76, wherein the cancer is refractory. 77. The method of claim 76, wherein the cancer recurs. 77. The method of claim 76, wherein the cancer is metastatic. 77. The method of claim 76, wherein said cancer is associated with overexpression of STAT3. 77. The method of claim 76, further comprising detecting a level of phosphorylated STAT3 (p-STAT3) in the tissue of the patient, wherein the level of p-STAT3 is used as a biomarker for patient selection. 77. The method of claim 76, wherein said cancer is associated with nuclear beta -catenin expression. 77. The method of claim 76, further comprising detecting beta -catenin expression in a tissue of a patient, wherein said beta -catenin expression is used as a biomarker for patient selection. 77. The method of claim 76, wherein significant beta -catenin expression is detected in the nucleus. CLAIMS We claim: 1. A method of treating cancer in a patient wherein expression of? -Catenin in the cell is detected in the nucleus, comprising administering to said patient a therapeutically effective amount of the following structure:

Or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof. CLAIMS 1. A method for treating cancer in a patient in which expression of beta -catenin in the cell is detected in the nucleus of the cell rather than in the cell membrane, comprising administering to said patient a therapeutically effective amount of the following structure:

Or a pharmaceutically acceptable salt, solvate, hydrate or prodrug thereof. 111. The method of claim 111, wherein the beta -catenin is present in intermediate to strong beta -catenin expression levels such that beta -catenin is detected in at least 20% tumor cells. (a) a therapeutically effective amount of the following structure:

&Lt; / RTI &gt;
(b) a polyoxyl glyceride having an HLB greater than 10; And
(c) a polyoxyl glyceride having an HLB of less than 10
&Lt; / RTI &gt; 114. The pharmaceutical composition of claim 113, further comprising a surfactant. 114. The pharmaceutical composition of claim 113, wherein the polyoxyl glyceride having an HLB greater than 10 is selected from the group consisting of lauroyl polyoxyl glyceride and stearoyl polyoxy glyceride. 114. The pharmaceutical composition of claim 113, wherein the polyoxyl glyceride having an HLB less than 10 is linoleoyl polyoxyl glyceride. 114. The pharmaceutical composition of claim 114, wherein the surfactant is sodium lauryl sulfate (SLS) or sodium dodecyl sulfate. 114. The method of claim 113 or 114 wherein the polyoxyl glyceride having an HLB of greater than 10 is selected from the group consisting of lauroyl polyoxyl glyceride and stearoyl polyoxy glyceride and the polyoxyl glyceride having an HLB less than 10 is selected from the group consisting of linoleoyl / RTI &gt; glycerides. 115. The method of Claim 114, wherein the polyoxyl glyceride having an HLB greater than 10 is selected from the group consisting of lauroyl polyoxyl glyceride and stearoyl polyoxyl glyceride, and the polyoxyl glyceride having an HLB less than 10 is linoleoyl polyoxyl glyceride , The surfactant is sodium lauryl sulfate (SLS) or sodium dodecyl sulfate.

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